Residual Brain Infection in Relapsing-Fever Borreliosis

Save this PDF as:
 WORD  PNG  TXT  JPG

Size: px
Start display at page:

Download "Residual Brain Infection in Relapsing-Fever Borreliosis"

Transcription

1 MAJOR ARTICLE Residual Brain Infection in Relapsing-Fever Borreliosis Diego Cadavid, 1,2 Marie Sondey, 1,2,a Edwin Garcia, 1,2,a and Catherine L. Lawson 3 1 Department of Neurology and Neuroscience and 2 Center for the Study of Emerging Pathogens, University of Medicine and Dentistry of New Jersey New Jersey Medical School, Newark, and 3 Department of Chemistry and Chemical Biology, Rutgers University, Piscataway, New Jersey Background. Neurological involvement is common in the spirochetal infection relapsing fever (RF) in both humans and experimental animals. RF is best known for antigenic variation caused by the sequential expression of variable outer membrane lipoproteins of 2 sizes, variable small (Vsp) and variable large (Vlp) proteins. Less understood is the persistence of RF borreliae in the brain after they are cleared from the blood, referred to as residual brain infection (RBI). Our goal was to investigate the phenomenon of RBI in RF. Methods. We studied RBI in immunocompetent mice by culturing blood and perfused brain samples 1 month after intraperitoneal inoculation with Borrelia turicatae serotype 1 (Bt1). Mice deficient in Toll-like receptor 2 (TLR2 / ) or in B and T cells (scid) were included for comparison. Results. All scid mice had persistent infection in blood and brain. RBI was found in 3 (19%) of 16 immunocompetent and TLR2 / mice. RBI was caused by either persistence of the original serotype (Bt1) or newly emerged Vsp ( n p 1, renamed Bt3) or Vlp serotypes. The Vsp of Bt1 (Vsp1) and Bt3 (Vsp3) were 75% identical. Conclusions. RBI in RF is relatively frequent and can occur by persistence of the original or newly emerged serotypes. Brain infection is a characteristic feature of many pathogenic spirochetes, notably Treponema palllidum and the several Borrelia species that cause Lyme disease (LD) and relapsing fever (RF). The neurological complications of Borrelia infections are collectively referred to as neuroborreliosis [1, 2]. The most common manifestations of neuroborreliosis in RF are meningitis, facial nerve palsy, radiculitis, and encephalopathy (reviewed in [3]). In outbreaks of tickborne RF, neuroborreliosis can be the dominant clinical feature [4, 5]. Our laboratory has been studying the pathogenesis of neuroborreliosis in animal models using nonhuman primates infected with LD borreliae and mice infected with RF borreliae [6 8]. RF borreliae are best known for the phenomenon of anti- Received 24 August 2005; accepted 9 December 2005; electronically published 4 April Potential conflicts of interest: none reported. Financial support: Foundation of the University of Medicine and Dentistry of New Jersey (grant to D.C.); Heritage Affiliate of the American Heart Association (Scientist Development Grant T to D.C.). a Present affiliations: Department of Biochemistry, University of Medicine and Dentistry of New Jersey New Jersey Medical School, Newark (M.S.); New York College of Osteopathic Medicine, Old Westbury (E.G.) Reprints or correspondence: Dr. Diego Cadavid, University of Medicine and Dentistry of New Jersey New Jersey Medical School, 185 S. Orange Ave., MSB H506, Newark, NJ The Journal of Infectious Diseases 2006; 193: by the Infectious Diseases Society of America. All rights reserved /2006/ $15.00 genic variation, which allows them to spontaneously change their serotype by switching the expression of variable major proteins (VMPs) [9, 10] of 2 sizes, large (Vlp) and small (Vsp). A switch in VMP allows RF borreliae to escape killing by the host s serotype-specific antibody response. Less known is the phenomenon of residual brain infection (RBI), which refers to the tendency of RF borreliae to persist in the brain after they disappear from the blood (reviewed in [1]). The goal of the present study was to investigate the phenomenon of RBI in RF. The results revealed that RBI is relatively frequent in both immunocompetent and Toll-like receptor 2 deficient (TLR2 / ) mice. RBI can be caused by the persistence of spirochetes in the brain, after their clearance from the blood, of either the original infecting serotype or new serotypes that spontaneously emerge during infection. MATERIALS AND METHODS Strains and culture conditions. B. turicatae serotypes 1 (Bt1; formerly serotype A) and 2 (Bt2; formerly serotype B) have been described elsewhere [7, 11 13]. Borreliae were cultured in BSK II medium [14] and counted in a Petroff-Hauser chamber [10]. When mouse tissue samples were cultured, 50 mg/ml rifampin and 100 mg/ ml phosphomycin were present in the medium. Plas- Residual Brain Infection in RF Borreliosis JID 2006:193 (15 May) 1451

2 Table 1. Susceptibility to neuroborreliosis in mice of different genetic background inoculated with serotype 1 of Borrelia turicatae. Mouse Blood Infected tissue Brain Serotype in brain a Vestibular dysfunction b Swiss Websters (outbred) 0/4 1/4 Novel 1/4 C3H/HeJ 0/4 1/4 Serotype 1 0/4 SWR/J 1/4 0/4 0/4 C57BL/6 0/4 1/4 Novel 0/4 C57BL/6 TLR2 / 4/16 3/16 All novel 0/16 C57BL/6 scid 4/4 4/4 Serotype 1 in all 4/4 C3H/HeJ scid 4/4 4/4 Serotype 1 in all 4/4 CB17 scid 4/4 4/4 Serotype 1 in all 4/4 NOTE. Data are no. of positive cultures/no. of tissue or fluid samples cultured. Mice were 3 inoculated intraperitoneally with 1 10 cells and infection examined by culture of blood or perfused brain 1 month later. TLR, Toll-like receptor. a By Western blot with anti variable small protein 1 polyclonal antibody. b Spinning in circles when lifted off the ground by the tail. ma samples from infected mice were used to start broth cultures, which were collected by centrifugation at 9000 g at cell densities of cells/ml. The purity of Bt1 was assessed before inoculation by Western blot with anti-vsp1 polyclonal antiserum [11, 12]. Mouse infections. Female mice, 4 6 weeks old, of different genetic backgrounds, purchased from Jackson or Charles Rivers or bred in-house (C57BL/6 TLR2 / ), were inoculated intraperitoneally with Bt1 spirochetes in 300 ml of PBS. Groups of 4 8 mice each were used for all experiments. Mice sham-inoculated with PBS were used as controls. The housing and care were in accordance with the Animal Welfare Act. Infection was confirmed by mixing tail-vein blood with an equal volume of PBS followed by dark-field microscopy. scid and C57BL/6-TLR2 / mice were maintained in a germ-free environment before and after infection. Tissue and fluid collection. Mice were anesthetized with isoflurane for euthanasia and necropsied as described elsewhere [6]. Total body perfusion with 30 ml of PBS was performed, to minimize blood contamination of the brain. Whole brains were homogenized in BSK II medium and centrifuged for 3 s at 7000 g; then, the supernatant inoculated into 6-mL BSK II culture tubes. Any brain homogenates in which blood contamination was detectable were discarded. All cultures from brain and blood were examined for spirochetes by phase-contrast microscopy for a 2-week period. Histopathological studies. Brains were removed at necropsy, fixed in 4% paraformaldehyde for 48 h at 4 C, and embedded in paraffin. Hematoxylin-eosin (HE) staining was prepared using standard techniques. All tissue sections were examined with standard light microscopy by a trained neuropathologist (D.C.) blinded to the infection status. Inflammation was defined in accordance with characteristic morphological changes after HE staining. A 3-step streptavidin-peroxidase technique was used for immunohistochemical analysis, as described elsewhere [15, 16]. Paraformaldehyde-fixed sections were treated with 0.5 mg/ml protease type VIII (Sigma P-5380) for 10 min for antigen retrieval. Commercially available rat monoclonal antibody anti mouse macrophage F4/80 at a 1:1000 dilution (Serotec) was used for the identification of activated macrophages/microglia. The chromogen was 3-3 -diaminobenzidine tetrahydrochloride. Tissue sections from uninfected mice were used as negative controls. Purified rat IgG of the same species (Sigma), matched for concentration and isotype, was used as a negative control. Image analysis was done by scoring microscopic fields per sagittal brain section as follows: 0, no F4/80-positive cells; 1, 1 2 cells/field; 2, 3 10 cells/field; 3, 110 cells/field; and 4, diffusely positive. The microscopic fields were from leptomeninges or brain parenchyma, and the examiner was blinded to infection status and genotype. Protein analysis. Whole cells from harvested cultures were subjected to SDS-PAGE analysis with 12.5% acrylamide [17]. For Western-blot analysis, proteins were transferred to nitrocellulose membranes (Millipore) that were then blocked with 3% (wt/vol) dried nonfat milk in 10 mmol/l Tris (ph 7.4) and 150 mmol/l NaCl (milk/ts) for 4 h [18]. After the membranes were washed 3 times with 0.3% milk/ts, they were incubated with anti-vsp1 polyclonal antiserum diluted 1:50,000 in 0.3% milk/ts [15]. Alkaline phosphatase conjugated anti rabbit antibody (Sigma) served as the second ligand. The blots were developed with nitroblue tetrazolium chloride/5-bromo 4-chloro 3-indolylphosphatase p-toluidine salt as the substrate (Pierce). DNA methods. Plasmid-rich Borrelia DNA was prepared using the diethylpyrocarbonate method [6]. All expressed vsp 1452 JID 2006:193 (15 May) Cadavid et al.

3 Table 2. Microglial/macrophage activation in brains of C57BL/6 wild-type or Toll-like receptor (TLR) 2 deficient (TLR2 / ) mice 1 month after inoculation of serotype 1 of Borrelia turicatae (Bt1) or PBS (as a control). Mouse strain, inoculation Brain parenchyma Leptomeninges C57BL/6 Bt (0 2.2) 7.5 ( ) PBS (0 1.7) C57BL/6 TLR2 / Bt ( ) PBS 0 0 NOTE. Macrophages and microglia were detected by immunostaining with rat anti-mouse F4/80 monoclonal antibody. Results are given as mean (95% confidence interval) sum score for microscopic fields. Scoring was done by an examiner who was blinded to infection status. The scoring system was as follows: 0, no F4/80-positive cells; 1, 1 2 cells/field; 2, 3 10 cells/field; 3, 110 cells/field; and 4, diffusely positive. genes were amplified and sequenced using a forward primer for the B. turicatae vsp promoter (5 -TTGAAAAATCGAAAT- TTTGAAATA-3, nt 9 32 on GenBank U85413) and a reversedegenerated primer for a conserved region on the 3 end of all known B. hermsii and B. turicatae vsp genes (5 -TGWAAGCK- SWKYDATTRCAG-3, nt on GenBank U854130) [11, 12]. When it is annealed at 42 C, this primer pair specifically amplifies expressed vsp and vlp genes from B. hermsii (Bh7 and Bh21) and B. turicatae (Bt1 and Bt2). Because only genes placed in front of the promoter are expressed, using a promoter specific primer allows the study of gene expression by polymerase chain reaction (PCR) instead of reverse-transcription PCR. DNA amplification was performed in a thermal cycler using 50-mL reaction volumes that contained 1.25 U of Thermus aquaticus DNA polymerase (Roche), 25 pmol of each primer, 200 mmol/ L each dntp, 10 mmol/l Tris (ph 8.3), 50 mmol/l KCl, and 4 mmol/l MgCl 2. The program was 40 cycles for 1 min at 94 C, for 1 min at 42 C, and for 2 min at 72 C, followed by a final extension cycle for 7 min at 72 C. Tubes with buffers and reagents but without DNA were included as negative controls for crosscontamination. DNA sequencing was performed 3 times on each strand using the cycle-sequencing method through commercial vendors (Retrogen). vsp3 was compared with previously characterized genes using WU-Blast2 software (version 2.0; Washington University). Vsp3 was compared to Vsp1 and Vsp2 using CLUSTAL W (version 1.82) multiple sequence alignment (available at: Statistical analysis. The 2-tailed Fisher s exact test was used to compare differences in percentages, and nonparametric tests were used to compare differences between means. P!.05 was considered to be significant. RESULTS RBI in immunocompetent mice. To study the susceptibility to RBI of immunocompetent mice, we examined outbred and inbred mice of different genetic backgrounds. All mice were inoculated intraperitoneally with Bt1 spirochetes, and the infection of blood and perfused brain was examined 1 month later by culture followed by SDS-PAGE and Westernblot analyses. The inbred strains were chosen arbitrarily on the basis of their known susceptibility to Lyme arthritis, another spirochetal disease. The strains chosen were C57BL/6 (H2b haplotype), which is moderately susceptible to Lyme arthritis [19], and C3H/HeJ (H2k) and SWR/J (H2q), which are highly susceptible [20]. Swiss Webster outbred mice were also included for the examination of RBI in a genetically heterogeneous population. RBI was defined as a positive brain culture in a mouse whose necropsy blood sample was culture negative. The results (table 1) showed that RBI was found in 3 (19%) of 16 immunocompetent mice. Although none of the inbred mice developed any signs of neurological dysfunction, 1 of 4 outbred mice developed severe head tilt, walking in circles, and spinning in the air when lifted off the ground by the tail. These manifestations of vestibular dysfunction were indistinguishable in time of onset and severity from those observed in scid mice (see below). Only 1 (6%) of 16 immunocompetent mice had positive blood cultures 1 month after inoculation (table 1). These results showed that RBI is relatively common in immunocompetent mice across different genetic backgrounds. Furthermore, they showed that, occasionally, immunocompetent mice develop vestibular dysfunction as a complication of RF borreliosis. Figure 1. Assay in 12.5% SDS-PAGE of brain (br) and blood (bl) culture lysates from 4 (1 4) C57BL/6-scid mice inoculated with serotype 1 of Borrelia turicatae (Bt1) and necropsied 1 month later. Molecular-weight markers are shown in kilodaltons on the left, and the whole serotype 1 spirochete lysate is shown on the right (Bt1), for comparison. The arrow points to 23-kDa variable small protein 1, the serotype-defining major outer membrane lipoprotein of Bt1. Residual Brain Infection in RF Borreliosis JID 2006:193 (15 May) 1453

4 RBI in TLR25/5 mice. The main virulence determinants of RF [21, 22] and LD [23] borreliae are lipoproteins. Because TLR2 is an important signal-transducing receptor for bacterial lipoproteins [24, 25], it is possible that TLR2 deficiency, by impairing recognition of the infection, might increase susceptibility to RBI in RF. To investigate this possibility, we cultured blood and perfused brain samples from TLR2 / mice 1 month after inoculation with Bt1. The results revealed RBI in 3 (19%) of 16 TLR2 / mice examined, similar to the frequency of persistent infection in blood samples (4/16 [25%]; table 1). Although the differences in persistent blood infection between immunocompetent (1/16 [6%]) and TLR2 / (4/16 [25%]) mice did not reach statistical significance, we cannot rule out a type II error, because the number of mice used was not large. It is also possible that TLR2 deficiency may influence the spirochetal load in other tissues that we did not study. None of the TLR2 / mice developed any signs of vestibular dysfunction. Because TLR2 plays an important role in the inflammatory response to LD borreliosis [25], we studied whether TLR2 deficiency had any effect in the inflammatory response to the infection in the brain of mice with RF borreliosis. For this, the brain from C57BL/6 wild-type and TLR2 / mice necropsied 1 month after intraperitoneal inoculation with Bt1 were examined microscopically. The examination of HE-stained slides showed minimal to mild meningitis in both groups (not shown). Immunohistochemical staining with an antibody to the glycoprotein F4/80, a marker of mouse microglia/macrophage activation [26], revealed increased signals, mainly in the leptomeninges. Image analysis of F4/80-immunostained sections revealed that the extent 1454 JID 2006:193 (15 May) Cadavid et al. Figure 3. Residual brain infection (RBI) in Toll-like receptor (TLR) 2 deficient (TLR2 / ) mice. A, SDS-PAGE; B, Western blot with anti variable small protein (Vsp1) rabbit polyclonal antibody of 3 TLR2 / mice showing RBI 1 month after intraperitoneal inoculation (lanes 2 4). Lane 1, Serotype 1 whole-cell lysate for comparison; lane 5, molecular-weight markers (in kilodaltons). B, Borrelia turicatae serotype 1 cell lysate but none of the brain culture lysates from TLR2 / mice reacting with anti-vsp1 antibody. Figure 2. Residual brain infection in immunocompetent mice. A, SDSPAGE and B, Western blot with anti variable small protein (Vsp1) polyclonal antibody of blood (lane 1) and brain (lanes 2 4) BSK II culture lysates from 4 immunocompetent mice (lanes 1 4) of different genetic backgrounds persistently infected with Borrelia turicatae (table 1). Molecular-weight markers are shown in kilodaltons on the left, and a lysate from serotype 1 (Bt1) is shown in lane 5 for comparison. B, Reactions only with the antivsp1 antibody (lanes 3 and 5). The lysate from B. turicatae serotype 2 spirochetes did not react with the anti-vsp1 antibody (data not shown). The arrow points to the 23-kDa Vsp1 in the Bt1-positive control. of macrophage/microglia activation was similar in wild-type and TLR2 / mice (table 2). These results revealed that TLR2 deficiency does not increase susceptibility to RBI or brain inflammation in RF borreliosis. Other factors can compensate for the lack of TLR2. RF neuroborreliosis in scid mice. Previous studies showed that CB17-scid mice are very susceptible to RF borreliosis, including brain infection, when they are inoculated with Bt1 [7, 16]. To investigate whether the susceptibility to brain infection in scid mice is influenced by other background genes, we compared neuroborreliosis in scid mice of 3 different genetic backgrounds CB17, C3H/HeJ, and C57BL/6 after inoculation with Bt1. In contrast to immunocompetent and TLR2 / mice, all scid mice developed persistent infection of the blood and brain and vestibular dysfunction, independently of their genetic background (table 1). SDS-PAGE analysis of positive blood and brain cultures from all scid mice showed the presence of a single VMP band of 23 kda (figure 1). As expected, Western-blot analysis with anti-vsp1 polyclonal antiserum confirmed that the band was Vsp1 (not shown). These results showed that the scid immunodeficiency significantly increases the frequency of blood and brain infection, compared with that in immunocompetent and TLR2 / mice, from just under 20% to 100% (P!.0001). Similarly, all scid mice developed clinical manifestations of vestibular dysfunction. Analysis of B. turicatae serotypes causing RBI. In contrast to the finding in scid mice, which all had a single VMP band of 23 kda (Vsp1), SDS-PAGE analysis of the 3 serotypes causing RBI in the immunocompetent mice showed that only 1 still had a 23-kDa VMP band (figure 2, lane 3). The other 2 (figure 2, lanes 2 and 4) had much larger VMP bands, corresponding to members of the Vlp families [27]. Similarly, the one serotype causing persistent blood infection in immunocompetent mice

5 Figure 4. Comparison of deduced protein sequences of variable small protein (Vsp) 1, Vsp2, and Vsp3 of Borrelia turicatae. CLUSTAL W (version 1.82) multiple sequence alignment (available at: showed that Vsp3 is very similar to Vsp1 and Vsp2 in the proximal and distal thirds, whereas the middle third is more heterogeneous. In the consensus line, an asterisk indicates strict conservation, meaning that the same residue is found in the considered position in compared amino acid sequences; a colon indicates conserved substitutions, meaning that chemically similar residues are found in the considered position in compared amino acid sequences (e.g., all basic residues [R or K], or all acidic residues [D or E]); and a period indicates semiconserved substitutions, meaning that residues found in the considered position are more variable than in the conserved case but are still considered to be chemically similar (e.g., all polar residues [R, K, D, E, S, or T]). (figure 2, lane 1) was also a Vlp member. Western-blot analysis with anti-vsp1 polyclonal antiserum revealed that the VMP band on lane 3 was Vsp1 (figure 2B). This was a brain culture from a C3H/HeJ mouse whose blood was found to be not infected at necropsy (table 1). This is, to our knowledge, the first time that RBI caused by the serotype originally inoculated has been demonstrated in RF. Analysis by SDS-PAGE of the 3 relapse serotypes causing RBI in TLR2 / mice (figure 3A, lanes 2 4) showed a similar situation: only 1 of the lanes (lane 3) had a VMP of 23 kda, identical to that of Vsp1 (lane 1). The other 2 (lanes 2 and 4) had VMP bands of 37 kda, a typical molecular size of members of the 5 Vlp families [28]. In contrast to immunocompetent mice, the 23-kDa VMP band in TLR2 / mice (figure 3B, lane 3) was not recognized by anti-vsp1 polyclonal antiserum. The implication of this finding is that a new member of the Vsp family of B. turicatae was responsible for RBI in 1 of the TLR2 / mice. This relapse serotype was cloned by limiting dilution in BSK II media [29], renamed serotype 3, and further characterized. Characterization of B. turicatae serotype 3. Serotype 3 spirochetes were found to be infectious for newly inoculated CB17-scid mice and were confirmed to be neurotropic by infecting the brain of all 3 scid mice examined (not shown). A PCR product of the predicted size was amplified from serotype 3 plasmid DNA using a forward primer specific for the B. turicatae vsp1 and vsp2 promoter region [11, 12] and a degenerated reverse primer based on a conserved sequence at the 3 end of all known vsp genes from B. hermsii and B. turicatae [27]. The deduced amino acid sequence (figure 4) revealed that Vsp3 is a member of the Vsp/OspC family of Borrelia proteins [30]. The proximal and distal thirds were highly conserved, whereas the middle third was hypervariable (figure 4). Analysis of the DNA sequence homology revealed the highest homology of vsp3 was with B. turicatae vsp1 (75%; table 3). vsp3 was 64% 74% similar to vmp genes from other RF borreliae and 62% 66% similar to ospc genes from LD borreliae (table 3). DISCUSSION The major findings of the present study were as follows. (1) RBI occurs in close to 20% of immunocompetent mice inoculated with a neurotropic strain of B. turicatae. (2) TLR2 deficiency does not increase susceptibility to RBI. (3) The scid mutation confers 100% susceptibility to RF neuroborreliosis, independently of background genes. (4) Vestibular dysfunction is the main clinical manifestation of RF neuroborreliosis in mice. (5) RBI can be caused by persistence of the original infecting serotype or by newly emerged serotypes that spontaneously arise in the population. Interest in the phenomenon of neurological involvement by Borrelia species dates to the early twentieth century, when neurological complications were frequent in patients with RF; however, research was almost abandoned after effective treatment with penicillin became available (reviewed in [1]). Earlier investigators in the field were especially interested in the ability of some RF spirochetes to persist in the brain for long time after they disappear from the blood, referred to as RBI [31, Residual Brain Infection in RF Borreliosis JID 2006:193 (15 May) 1455

6 Table 3. Genes most closely related to Borrelia turicatae vsp3. Borrelia species Disease Genes Similarity, a % B. turicatae Relapsing fever in the US vsp1 b 75 vsp2, vsp4, vsp5, vsp6 74 B. hermsii Relapsing fever in the US vsp3, vlp7, vlp21 68 vsp3, vsp22, vsp13, vsp26 67 vsp24, vsp6, vsp17, vsp25 66 B. duttoni Relapsing fever in Africa vmp (unnamed) 66 B. valaisiana Lyme disease like species from Eurasia c ospc 66 B. afzelii Lyme disease in Europe ospc 65 B. miyamotoi Relapsing fever like species from Japan c vmp (unnamed) 64 B. burgdorferi sensu stricto Lyme disease in the Western Hemisphere B. garinii Lyme disease in Europe ospc 62 ospc 62 a Determined using the Washington University Basic Local Alignment Search Tool (version 2.0). b The nomenclature for the variable major protein family of relapsing-fever borrelia genes has been changed from the original vmp to vsp (variable small protein) or vlp (variable large protein) genes, and letters have been replaced by nos. For example, Vsp1 was initially referred to as VmpA [11], then VspA [49]. c These species have been isolated from ticks and rodents, but no human infections have been reported. 32]. RBI was the subject of many investigations during the last century [1]. It was shown to occur not only in experimental animals but also in nature [33, 34]. RBI was documented in experimental animals for up to 3 years after inoculation [35]. Borreliae causing RBI are susceptible to the serum from the animal from which they are recovered, and they cannot reenter the blood from the brain [36]. In contrast to RF, RBI has not been documented in animal models of Lyme borreliosis [37 39]. In a recent study, we did not find a single case of RBI in 40 outbred mice 1 month after inoculation with several different strains of LD borreliae, most of which were isolated from human cerebrospinal fluid [40]. In contrast, brain involvement is a frequent occurrence with many species of RF borreliae [1, 6, 7, 41, 42]. A major recent advance in the field of RF has been the elucidation by Barbour et al. [43 45] that a switch in the expression of VMP proteins due to spontaneous gene conversion of silent vmp genes to a single expression locus in a linear plasmid is the underlying mechanism of antigenic variation in RF. However, whether the VMP switch influences brain infection remains to be determined. The results reported in the present article reveal that the main determinant of susceptibility to brain infection in RF is the ability to mount a specific immune response. This is in agreement with previous studies that showed that the ability to mount a specific antibody response is the main determinant of susceptibility to RF [6, 7]. Recent studies have shown that these antibodies are produced by B1b lymphocytes [46] and that they can be bactericidal in the absence of complement [47]. Although scid mice are deficient in both B and T cells, B cell deficiency is likely responsible for their increased susceptibility to RF borreliosis [48]. In support of this is our recent finding that Igh6- deficient mice, which are deficient only in B cells, are also highly susceptible to neuroborreliosis after inoculation with Bt1 (D.C. and H. Gelderblom, data not shown). In contrast to the scid mutation, we did not find any evidence that TLR2 deficiency increases susceptibility to RF neuroborreliosis: the frequency of RBI, cerebral microgliosis, and vestibular dysfunction was similar in wild-type and TLR2-deficient mice. This is in contrast to the case in Lyme borreliosis, in which TLR2 deficiency increases the severity of arthritis and the spirochetal load in some tissues [25]. Previous studies of scid mice inoculated with B. turicatae [7, 15] and Balb/c mice inoculated with Borrelia hermsii [6] showed significant differences in the ability of the individual serotypes to enter the brain. Because the only difference between isogenic Borrelia serotypes is their VMPs, this suggests that expressing certain VMPs (like Vsp1) may facilitate brain invasion. In other words, some RF VMPs may function as spirochetal neuroinvasins. This suggests that another consequence of VMP variation in RF may be a gain or loss of function with regard to brain infection. Sequence analysis of Vsp3 showed that it is closely related to Vsp1 (table 3 and figure 4). This suggests that a VMP switch from Vsp1 to Vsp3 may have allowed B. turicatae to escape killing while still preserving its ability to infect the brain. Sequence alignment of B. turicatae Vsp1, 2, and 3 showed they are very similar in the proximal and distal regions, with most of the variability concentrated in the middle third (figure 4). Localized polymorphisms in this middle region of Vsp [49] may account for differences of individual Borrelia serotypes in the ability to infect the brain. In previous studies of Balb/c mice inoculated with B. hermsii serotype 7, we found that it had been cleared not only from the blood but also from the brain by the time of the first relapse [6]. In this study, we also found that an established serotype 7 brain infection was cleared from the brain of irradiated Balb/ 1456 JID 2006:193 (15 May) Cadavid et al.

7 c mice by the intravenous administration of anti-vsp7 monoclonal antibodies of the IgG but not of the IgM isotype [6]. This demonstrated that some antibodies are better than others at clearing spirochetes from the brain. However, we have now found Bt1 in the brain of an immunocompetent mouse that had been inoculated with Bt1 1 month earlier. This is, to our knowledge, the first demonstration that the original infecting serotype can somehow escape the host s antibody response and survive in the brain. The implication is that, occasionally, immunocompetent hosts fail to clear spirochetes from the brain, resulting in RBI. The mechanism that allowed Bt1 to survive in the brain but not in the blood remains to be determined. Because the spirochetes cultured from the brain were expressing Vsp1, it is more likely that the antibody response of this particular C3H/HeJ mouse had difficulty reaching or killing the spirochetes that had entered the brain. Although the number of mice was small, it was also interesting that one of the outbred mice developed vestibular dysfunction as severe as that of scid mice. This also implies that some hosts, who apparently are immunocompetent, fail to control RF borreliosis, resulting in neurological complications. Contrary to the case in humans, in whom neuroborreliosis involves predominantly the seventh cranial nerve both in RF and LD [3], in mice, RF neuroborreliosis predominantly affects the vestibular system [7, 15]. The present results reveal that persistent brain infection in RF borreliosis is not only a feature of antibody-deficient mice but that it also occurs in a significant percentage of immunocompetent mice. RBI can be caused by the persistence in the brain of the original or relapse serotypes after they had been eliminated from the blood. Future studies may elucidate how variations in VMPs modulate the ability of RF borreliae to enter the brain. Acknowledgment We thank Dr. Shizuo Akira (Osaka University, Japan), for his gift of the Toll-like receptor 2 deficient mice. References 1. Cadavid D, Barbour AG. Neuroborreliosis during relapsing fever: review of the clinical manifestations, pathology, and treatment of infections in humans and experimental animals. Clin Infect Dis 1998; 26: Pachner AR, Cadavid D. Lyme neuroborreliosis. In: Griggs RC, Joynt RJ, eds. Clinical neurology. Philadelphia: Lippincott-Raven, 1998: Cadavid D. Lyme disease and relapsing fever. In: Scheld W, Whitley R, Marra C, eds. Infections of the central nervous system. Philadelphia: Lippincott-Raven, 2004: Taft W, Pike J. Relapsing fever: report of a sporadic outbreak including treatment with penicillin. JAMA 1945; 129: Bergeret C, Raoult A. Notes sur les formes nerveuses de la fievre recurrente fievre recurrente a tiques en Afrique Occidentale Francaise. Bulletin Medicale de l Afrique Occidentale Francaise 1948; 5: Cadavid D, Bundoc V, Barbour AG. Experimental infection of the mouse brain by a relapsing fever Borrelia species: a molecular analysis. J Infect Dis 1993; 168: Cadavid D, Thomas DD, Crawley R, Barbour AG. Variability of a bacterial surface protein and disease expression in a possible mouse model of systemic Lyme borreliosis. J Exp Med 1994; 179: Cadavid D, O Neill T, Schaefer H, Pachner AR. Localization of Borrelia burgdorferi in the nervous system and other organs in a nonhuman primate model of Lyme disease. Lab Invest 2000; 80: Plasterk RH, Simon MI, Barbour AG. Transposition of structural genes to an expression sequence on a linear plasmid causes antigenic variation in the bacterium Borrelia hermsii. Nature 1985; 318: Stoenner H, Dodd T, Larsen C. Antigenic variation of Borrelia hermsii. J Exp Med 1982; 156: Cadavid D, Pennington PM, Kerentseva TA, Bergstrom S, Barbour AG. Immunologic and genetic analyses of VmpA of a neurotropic strain of Borrelia turicatae. Infect Immun 1997; 65: Pennington PM, Cadavid D, Barbour AG. Characterization of VspB of Borrelia turicatae, a major outer membrane protein expressed in blood and tissues of mice. Infect Immun 1999; 67: Pennington PM, Cadavid D, Bunikis J, Norris SJ, Barbour AG. Extensive interplasmidic duplications change the virulence phenotype of the relapsing fever agent Borrelia turicatae. Mol Microbiol 1999; 34: Barbour AG, Hayes SF. Biology of Borrelia species. Microbiol Rev 1986; 50: Cadavid D, Pachner AR, Estanislao L, Patalapati R, Barbour AG. Isogenic serotypes of Borrelia turicatae show different localization in the brain and skin of mice. Infect Immun 2001; 69: Londoño D, Bai Y, Zuckert W, Gelderbrom H, Cadavid D. Cardiac apoptosis in severe relapsing fever borreliosis. Infect Immun 2005; 76: Barbour AG, Tessier SL, Stoenner HG. Variable major proteins of Borrelia hermsii. J Exp Med 1982; 156: Sadziene A, Rosa PA, Thompson PA, Hogan DM, Barbour AG. Antibody-resistant mutants of Borrelia burgdorferi: in vitro selection and characterization. J Exp Med 1992; 176: Schaible UE, Kramer MD, Wallich R, Tran T, Simon MM. Experimental Borrelia burgdorferi infection in inbred mouse strains: antibody response and association of H-2 genes with resistance and susceptibility to development of arthritis. Eur J Immunol 1991; 21: Barthold SW, Beck DS, Hansen GM, Terwilliger GA, Moody KD. Lyme borreliosis in selected strains and ages of laboratory mice. J Infect Dis 1990; 162: Barstad PA, Coligan JE, Raum MG, Barbour AG. Variable major proteins of Borrelia hermsii: epitope mapping and partial sequence analysis of CNBr peptides. J Exp Med 1985; 161: Vidal V, Scragg IG, Cutler SJ, et al. Variable major lipoprotein is a principal TNF-inducing factor of louse-borne relapsing fever. Nat Med 1998; 4: Fraser CM, Casjens S, Huang WM, et al. Genomic sequence of a Lyme disease spirochaete, Borrelia burgdorferi. Nature 1997; 390: Lien E, Sellati TJ, Yoshimura A, et al. Toll-like receptor 2 functions as a pattern recognition receptor for diverse bacterial products. J Biol Chem 1999; 274: Wooten RM, Ma Y, Yoder RA, et al. Toll-like receptor 2 is required for innate, but not acquired, host defense to Borrelia burgdorferi. J Immunol 2002; 168: Perry VH, Hume DA, Gordon S. Immunohistochemical localization of macrophages and microglia in the adult and developing mouse brain. Neuroscience 1985; 15: Hinnebusch BJ, Barbour AG, Restrepo BI, Schwan TG. Population structure of the relapsing fever spirochete Borrelia hermsii as indicated by polymorphism of two multigene families that encode immunogenic outer surface lipoproteins. Infect Immun 1998; 66: Burman N, Bergstrom S, Restrepo BI, Barbour AG. The variable antigens Vmp7 and Vmp21 of the relapsing fever bacterium Borrelia Residual Brain Infection in RF Borreliosis JID 2006:193 (15 May) 1457

8 hermsii are structurally analogous to the VSG proteins of the African trypanosome. Mol Microbiol 1990; 4: Barbour A. Isolation and cultivation of Lyme disease spirochetes. Yale J Biol Med 1984; 57: Carter CJ, Bergstrom S, Norris SJ, Barbour AG. A family of surfaceexposed proteins of 20 kilodaltons in the genus Borrelia. Infect Immun 1994; 62: Rothermundt M. Ueber die beziehungen zwischen virulenz und persistenz der rekurrensspirochaten im gehirn weisser mause. Arbeiten aus dem Staatsins fur Experimentelle Therapie und dem Georg-Speyer-Haus zu Frankfurt AM 1928: Levaditi C, Anderson T, Selbie F, Schoen R. L encephalite recurrentielle. Bull Acad Med 1930; 103: Diatta G, Trape J, Legros F, Rogier C, Duplantier J. A comparative study of three methods of detectin of Borrelia crocidurae in wild rodents in Senegal. Trans R Soc Trop Med Hyg 1994; 88: Horrenberger R. Spirochaeta hispanica chez les rats d Alger (nouvelle enquete). Arch Inst Pasteur Alger 1954; 32: Sergent E. Persistence de spirochaete hispanica pendant trois ans dans le cerveau d un cobaye. 3e note. Arch Inst Pasteur Alger 1945; 23: Steiner G, Steinfeld J, Schauder H. Zur frage der spirochatenpersistenzim zentralnervensystem bei experimenteller recurrens. Klinische Wochenschrift 1925; 4: Schaible UE, Gay S, Museteanu C, et al. Lyme borreliosis in the severe combined immunodeficiency (scid) mouse manifests predominantly in the joints, heart, and liver. Am J Pathol 1990; 137: Barthold SW, de Souza MS, Janotka JL, Smith AL, Persing DH. Chronic lyme borreliosis in the laboratory mouse. Am J Pathol 1993; 143: Pachner AR, Itano A. Borrelia burgdorferi infection of the brain: characterization of the organism and response to antibiotics and immune sera in the mouse model. Neurology 1990; 40: Pachner AR, Dail D, Bai Y, et al. Genotype determines phenotype in experimental Lyme neuroborreliosis. Ann Neurol 2004; 56: Nordstrand A, Shamaei-Tousi A, Ny A, Bergstrom S. Delayed invasion of the kidney and brain by Borrelia crocidurae in plasminogen-deficient mice. Infect Immun 2001; 69: Gebbia JA, Monco JC, Degen JL, Bugge TH, Benach JL. The plasminogen activation system enhances brain and heart invasion in murine relapsing fever borreliosis. J Clin Invest 1999; 103: Meier J, Simon M, Barbour A. Antigenic variation is associated with DNA rearrangements in a relapsing fever borrelia. Cell 1985; 41: Barbour A. Antigenic variation of a relapsing fever Borrelia species. Annu Rev Microbiol 1990; 44: Barbour AG, Burman N, Carter CJ, Kitten T, Bergstrom S. Variable antigen genes of the relapsing fever agent Borrelia hermsii are activated by promoter addition. Mol Microbiol 1991; 5: Alugupalli KR, Leong JM, Woodland RT, Muramatsu M, Honjo T, Gerstein RM. B1b lymphocytes confer T cell-independent long-lasting immunity. Immunity 2004; 21: Connolly SE, Thanassi DG, Benach JL. Generation of a complementindependent bactericidal IgM against a relapsing fever Borrelia. J Immunol 2004; 172: Newman K Jr, Johnson RC. T-cell-independent elimination of Borrelia turicatae. Infect Immun 1984; 45: Zuckert WR, Kerentseva TA, Lawson CL, Barbour AG. Structural conservation of neurotropism-associated VspA within the variable Borrelia Vsp-OspC lipoprotein family. J Biol Chem 2001; 276: JID 2006:193 (15 May) Cadavid et al.

16S rdna Sequencing Diagnosis of Spirochetemia in Lyme and related Borrelioses

16S rdna Sequencing Diagnosis of Spirochetemia in Lyme and related Borrelioses 16S rdna Sequencing Diagnosis of Spirochetemia in Lyme and related Borrelioses Sin Hang Lee, MD, F.R.C.P.(C)* Pathologist, Milford Hospital and Director, Milford Medical Laboratory, Inc. Milford, CT Email:

More information

Inactivation of Genes for Antigenic Variation in the Relapsing Fever Spirochete Borrelia hermsii Reduces Infectivity in Mice and Transmission by Ticks

Inactivation of Genes for Antigenic Variation in the Relapsing Fever Spirochete Borrelia hermsii Reduces Infectivity in Mice and Transmission by Ticks University of Montana ScholarWorks at University of Montana Biological Sciences Faculty Publications Biological Sciences 4-3-2014 Inactivation of Genes for Antigenic Variation in the Relapsing Fever Spirochete

More information

1 Name. 1. (3 pts) What is apoptosis and how does it differ from necrosis? Which is more likely to trigger inflammation?

1 Name. 1. (3 pts) What is apoptosis and how does it differ from necrosis? Which is more likely to trigger inflammation? 1 Name MCB 150 Midterm Eam #1 (100 points total) Please write your full name on each page of the eam!! The eam consists of 17 questions (6 pages). Each has a different point count as indicated. Please

More information

SOX2 antibody - Embryonic Stem Cell Marker (ab15830) datasheet

SOX2 antibody - Embryonic Stem Cell Marker (ab15830) datasheet -Embryonic Stem Cell Marker (ab15830) 1/8 ページ d Products: Stem Cells >> Germline Stem Cells >> Embryonic Germ Cells SOX2 antibody - Embryonic Stem Cell Marker (ab15830) datasheet Product Name Product type

More information

SANTA CRUZ BIOTECHNOLOGY, INC.

SANTA CRUZ BIOTECHNOLOGY, INC. TECHNICAL SERVICE GUIDE: Western Blotting 2. What size bands were expected and what size bands were detected? 3. Was the blot blank or was a dark background or non-specific bands seen? 4. Did this same

More information

Supplementary Material. Manuscript title: Cross-immunity and community structure of a multiple-strain pathogen in the

Supplementary Material. Manuscript title: Cross-immunity and community structure of a multiple-strain pathogen in the Supplementary Material Manuscript title: Cross-immunity and community structure of a multiple-strain pathogen in the tick vector Description of the primers used in the second PCR: The forward and the reverse

More information

WesternMAX Alkaline Phosphatase Chemiluminescent Detection Kits

WesternMAX Alkaline Phosphatase Chemiluminescent Detection Kits WesternMAX Alkaline Phosphatase Chemiluminescent Detection Kits Code N221-KIT N220-KIT Description WesternMAX Chemiluminescent AP Kit, Anti-Mouse Includes: Alkaline Phosphatase (AP) Conjugated Anti-Mouse

More information

Cell & Tissue Staining Kit

Cell & Tissue Staining Kit Cell & Tissue Staining Kit For the detection of goat, mouse, rabbit, rat, or sheep primary IgG Antibodies Size: 50 Tests HRP-DAB System Goat Kit (Catalog Number CTS008) Mouse Kit (Catalog Number CTS002)

More information

Effect of Immunization with Recombinant OspA on Serologic Tests for Lyme Borreliosis

Effect of Immunization with Recombinant OspA on Serologic Tests for Lyme Borreliosis CLINICAL AND DIAGNOSTIC LABORATORY IMMUNOLOGY, Jan. 2001, p. 79 84 Vol. 8, No. 1 1071-412X/01/$04.00 0 DOI: 10.1128/CDLI.8.1.79 84.2001 Copyright 2001, American Society for Microbiology. All Rights Reserved.

More information

Immunoprecipitation Protocol

Immunoprecipitation Protocol Immunoprecipitation Protocol Immunoprecipitation is a general method to obtain the enrichment of a specific protein from tissue lysate and cell lysate. It can be used to purify a specific protein, to identify

More information

Rat IGF-1 ELISA Kit (rigf-1-elisa)

Rat IGF-1 ELISA Kit (rigf-1-elisa) Rat IGF-1 ELISA Kit (rigf-1-elisa) Cat. No. EK0377 96 Tests in 8 x 12 divisible strips Background Insulin-like growth factor 1 (IGF-1), also known as somatomedin C, is a polypeptide protein hormone similar

More information

Technical Note Detection of post-immunoprecipitation proteins by Western blot using the Quick Western Kit IRDye 680RD

Technical Note Detection of post-immunoprecipitation proteins by Western blot using the Quick Western Kit IRDye 680RD Technical Note Detection of post-immunoprecipitation proteins by Western blot using the Quick Western Kit IRDye 680RD Developed for: Aerius, Odyssey Classic, Odyssey CLx and Odyssey Sa Imaging Systems

More information

Store samples to be assayed within 24 hours at 2-8 C. For long-term storage, aliquot and freeze samples at -20 C. Avoid repeated freeze-thaw cycles.

Store samples to be assayed within 24 hours at 2-8 C. For long-term storage, aliquot and freeze samples at -20 C. Avoid repeated freeze-thaw cycles. Human Retinol Binding Protein 4, RBP4 ELISA Kit Preparation Plate Washing Discard the solution in the plate without touching the side walls. Blot the plate onto paper towels or other absorbent material.

More information

Characterization of a Tick Isolate of Borrelia burgdorferi That

Characterization of a Tick Isolate of Borrelia burgdorferi That JOURNAL OF CLINICAL MICROBIOLOGY, June 1990, p. 1362-1366 0095-1137/90/061362-05$02.00/0 Copyright C 1990, American Society for Microbiology Vol. 28, No. 6 Characterization of a Tick Isolate of Borrelia

More information

The Expression of Recombinant Sheep Prion Protein (RecShPrPC) and its Detection Using Western Blot and Immuno-PCR

The Expression of Recombinant Sheep Prion Protein (RecShPrPC) and its Detection Using Western Blot and Immuno-PCR The Expression of Recombinant Sheep Prion Protein (RecShPrPC) and its Detection Using Western Blot and Immuno-PCR S. Thomas, C. S. Fernando, J. Roach, U. DeSilva and C. A. Mireles DeWitt The objective

More information

LAMININ. For Immunohistochemical Demonstration of Laminin in Paraffin-embedded and Frozen Human Tissue Sections Stock No. IMMH-7

LAMININ. For Immunohistochemical Demonstration of Laminin in Paraffin-embedded and Frozen Human Tissue Sections Stock No. IMMH-7 LAMININ For Immunohistochemical Demonstration of Laminin in Paraffin-embedded and Frozen Human Tissue Sections Stock No. IMMH-7 TABLE OF CONTENTS BACKGROUND AND PRINCIPLE... 4 REAGENTS AND EQUIPMENT PROVIDED...

More information

Suppression of Polyclonal Immunoglobulin Production by M-proteins Shows Isotype Specificity

Suppression of Polyclonal Immunoglobulin Production by M-proteins Shows Isotype Specificity 274 Annals of Clinical & Laboratory Science, vol. 31, no. 3, 2001 Suppression of Polyclonal Immunoglobulin Production by M-proteins Shows Isotype Specificity Liang Wang and David C. Young Department of

More information

High Pure PCR Template Preparation Kit for preparation of 100 nucleic acid samples Cat. No

High Pure PCR Template Preparation Kit for preparation of 100 nucleic acid samples Cat. No for preparation of 100 nucleic acid samples Cat. No. 1 796 88 Principle Cells are lysed during a short incubation with Proteinase K in the presence of a chaotropic salt (guanidine HCl), which immediately

More information

Manufactured by. Zyagen Barnes Canyon Road San Diego, CA 92121, USA

Manufactured by. Zyagen Barnes Canyon Road San Diego, CA 92121, USA Alkaline Phosphatase Immunohistochemistry Detection kits For detection of mouse, rabbit, goat, rat, sheep, chicken, guinea pig, and human primary antibodies Size: 500 Tests Catalog #: AK-011, Mouse Kit

More information

Segments of the obstructed intestinal loops were fixed in 4% paraformaldehyde

Segments of the obstructed intestinal loops were fixed in 4% paraformaldehyde Supplementary text Supplementary materials and methods Histopathological examination Segments of the obstructed intestinal loops were fixed in 4% paraformaldehyde (PFA) and embedded in paraffin wax with

More information

Borrelia burgdorferi Sensu Lato Species in Europe Induce Diverse Immune Responses against C 6 Peptides in Infected Mice

Borrelia burgdorferi Sensu Lato Species in Europe Induce Diverse Immune Responses against C 6 Peptides in Infected Mice CLINICAL AND VACCINE IMMUNOLOGY, Nov. 2009, p. 1546 1562 Vol. 16, No. 11 1556-6811/09/$12.00 doi:10.1128/cvi.00201-09 Copyright 2009, American Society for Microbiology. All Rights Reserved. Borrelia burgdorferi

More information

The majority of bacteria exist in nature attached to a substratum MacEachran, D.P. O Toole, G.A. The Biofilm Mode of Life, 2007 p23.

The majority of bacteria exist in nature attached to a substratum MacEachran, D.P. O Toole, G.A. The Biofilm Mode of Life, 2007 p23. The majority of bacteria exist in nature attached to a substratum MacEachran, D.P. O Toole, G.A. The Biofilm Mode of Life, 2007 p23. Biofilms of Borrelia burgdorferi And Clinical Implications for Chronic

More information

Updated April 27, PRODUCT INSERT

Updated April 27, PRODUCT INSERT Updated April 27, 2009 1 PRODUCT INSERT Hypoxyprobe, Inc. 121 Middlesex Turnpike Burlington, MA 01803 USA www.hypoxyprobe.com Hypoxyprobe Gemini Kit Kit Contents: Solid pimonidazole HCl (Hypoxyprobe -1)

More information

Human Rheumatoid Factor IgM ELISA Kit

Human Rheumatoid Factor IgM ELISA Kit Product Manual Human Rheumatoid Factor IgM ELISA Kit Catalog Number PRB- 5066 PRB- 5066 96 assays 5 x 96 assays FOR RESEARCH USE ONLY Not for use in diagnostic procedures Introduction Rheumatoid arthritis

More information

The GeneEditor TM in vitro Mutagenesis System: Site- Directed Mutagenesis Using Altered Beta-Lactamase Specificity

The GeneEditor TM in vitro Mutagenesis System: Site- Directed Mutagenesis Using Altered Beta-Lactamase Specificity Promega Notes Magazine Number 62, 1997, p. 02 The GeneEditor TM in vitro Mutagenesis System: Site- Directed Mutagenesis Using Altered Beta-Lactamase Specificity By Christine Andrews and Scott Lesley Promega

More information

Chapter 17: Immunization & Immune Testing. 1. Immunization 2. Diagnostic Immunology

Chapter 17: Immunization & Immune Testing. 1. Immunization 2. Diagnostic Immunology Chapter 17: Immunization & Immune Testing 1. Immunization 2. Diagnostic Immunology 1. Immunization Chapter Reading pp. 505-511 What is Immunization? A method of inducing artificial immunity by exposing

More information

Practical Applications of Immunology (Chapter 18) Lecture Materials for Amy Warenda Czura, Ph.D. Suffolk County Community College Eastern Campus

Practical Applications of Immunology (Chapter 18) Lecture Materials for Amy Warenda Czura, Ph.D. Suffolk County Community College Eastern Campus Practical Applications of Immunology (Chapter 18) Lecture Materials for Amy Warenda Czura, Ph.D. Suffolk County Community College Eastern Campus Primary Source for figures and content: Tortora, G.J. Microbiology

More information

MATF Antigen Submission Details and Standard Project Deliverables

MATF Antigen Submission Details and Standard Project Deliverables MATF Antigen Submission Details and Standard Project Deliverables What we require when you submit your antigen: Proteins For a recombinant protein target, we require a minimum of 400µg soluble recombinant

More information

Custom Antibodies Services. GeneCust Europe. GeneCust Europe

Custom Antibodies Services. GeneCust Europe. GeneCust Europe GeneCust Europe Laboratoire de Biotechnologie du Luxembourg S.A. 2 route de Remich L-5690 Ellange Luxembourg Tél. : +352 27620411 Fax : +352 27620412 Email : info@genecust.com Web : www.genecust.com Custom

More information

Reagents and cell culture Mcl-1 gene expression: real-time quantitative RT-PCR In vitro PP2A phosphatase assay Detection of Mcl-1 in vivo

Reagents and cell culture Mcl-1 gene expression: real-time quantitative RT-PCR In vitro PP2A phosphatase assay Detection of Mcl-1 in vivo Reagents and cell culture Antibodies specific for caspase 3, PARP and GAPDH were purchased from Cell Signaling Technology Inc. (Beverly, MA). Caspase inhibitor z-vad-fmk and ROS scavenger N-acetyl-Lcysteine

More information

obtained from the infected and treated tissues, Fleming's2 technic of hemolytic streptococcus B. Immediately following the infection, 1.0 ml.

obtained from the infected and treated tissues, Fleming's2 technic of hemolytic streptococcus B. Immediately following the infection, 1.0 ml. THE SENSITIVITY OF STREPTOCOCCI TO PENICILLIN G AFTER EXPOSURE TO THE ANTIBIOTIC IN VIVO* E. GRUNBERG, C. UNGER, AND D. ELDRIDGE Previous investigations by Grunberg, Schnitzer, and Unger3 on the topical

More information

Human Fibrinogen Antigen Assay

Human Fibrinogen Antigen Assay Catalog # FIBKT 0 Human Fibrinogen Antigen Assay Strip well format. Reagents for up to 96 tests. For Research Use Only. INTENDED USE This human fibrinogen antigen assay is intended for the quantitative

More information

Research Biochemicals. ANTIBODIES TO PRION PROTEINS

Research Biochemicals. ANTIBODIES TO PRION PROTEINS Research Biochemicals. ANTIBODIES TO PRION PROTEINS 2005_03 Research Biochemicals. TABLE OF CONTENTS MONOCLONAL AB 6H4 3 MONOCLONAL AB 34C9 4 RECOMBINANT BOVINE PRP 5 PRICE LIST 7 ORDER INFORMATION 8 2

More information

Discovery and Humanization of Novel High Affinity Neutralizing Monoclonal Antibodies to Human IL-17A

Discovery and Humanization of Novel High Affinity Neutralizing Monoclonal Antibodies to Human IL-17A Discovery and Humanization of Novel High Affinity Neutralizing Monoclonal Antibodies to Human IL-17A Contacts: Marty Simonetti martysimonetti@gmail.com Kirby Alton kirby.alton@abeomecorp.com Rick Shimkets

More information

Strategies for Assessment of Immunotoxicology in Preclinical Drug Development

Strategies for Assessment of Immunotoxicology in Preclinical Drug Development Strategies for Assessment of Immunotoxicology in Preclinical Drug Development Rebecca Brunette, PhD Scientist, Analytical Biology SNBL USA Preclinical Immunotoxicology The study of evaluating adverse effects

More information

Week 1: Protein isolation and quantification

Week 1: Protein isolation and quantification Week 1: Protein isolation and quantification Objective The objective of this lab exercise is to obtain protein samples from fruit fly larvae, BCS and FBS, all of which are then quantitated in the preparation

More information

Immunoglobulins. Harper s biochemistry Chapter 49

Immunoglobulins. Harper s biochemistry Chapter 49 Immunoglobulins Harper s biochemistry Chapter 49 Immune system Detects and inactivates foreign molecules, viruses, bacteria and microorganisms Two components with 2 strategies B Lymphocytes (humoral immune

More information

EAE Induction by Active Immunization in C57BL/6 Mice

EAE Induction by Active Immunization in C57BL/6 Mice Recommended protocol for use with Hooke Kit MOG 35-55/CFA Emulsion PTX (EK-0110, EK-0112, EK-0113, EK-0114, or EK-0115). Summary C57BL/6 mice develop chronic paralysis after immunization with MOG 35-55

More information

Recombinant DNA Technology

Recombinant DNA Technology History of recombinant DNA technology Recombinant DNA Technology (DNA cloning) Majid Mojarrad Recombinant DNA technology is one of the recent advances in biotechnology, which was developed by two scientists

More information

Antibody Structure. Antibodies

Antibody Structure. Antibodies Antibodies Secreted by B lymphocytes Great diversity and specificity: >10 9 different antibodies; can distinguish between very similar molecules Tag particles for clearance/destruction Protect against

More information

Antibody Structure supports Function

Antibody Structure supports Function Antibodies Secreted by B lymphocytes Great diversity and specificity: >10 9 different antibodies; can distinguish between very similar molecules Tag particles for clearance/destruction Protect against

More information

1. Cross-linking and cell harvesting

1. Cross-linking and cell harvesting ChIP is a powerful tool that allows the specific matching of proteins or histone modifications to regions of the genome. Chromatin is isolated and antibodies to the antigen of interest are used to determine

More information

Blot: a spot or stain, especially of ink on paper.

Blot: a spot or stain, especially of ink on paper. Blotting technique Blot: a spot or stain, especially of ink on paper. 2/27 In molecular biology and genetics, a blot is a method of transferring proteins, DNA or RNA, onto a carrier (for example, a nitrocellulose,pvdf

More information

Effects of Antibiotic Treatment on the Results of Nested ACCEPTED. Polymerase Chain Reactions for Scrub Typhus

Effects of Antibiotic Treatment on the Results of Nested ACCEPTED. Polymerase Chain Reactions for Scrub Typhus JCM Accepts, published online ahead of print on 0 August 00 J. Clin. Microbiol. doi:0./jcm.00-0 Copyright 00, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

More information

For the quantitative detection of human IL6 in serum, plasma, cell culture supernatants and urine.

For the quantitative detection of human IL6 in serum, plasma, cell culture supernatants and urine. m andw da a For the quantitative detection of human IL6 in serum, plasma, cell culture supernatants and urine. general information Catalogue Number Product Name Species cross-reactivity Range (calibration

More information

Mouse Monoclonal Antibody Isotyping Reagents

Mouse Monoclonal Antibody Isotyping Reagents Mouse Monoclonal Antibody Isotyping Reagents Catalog Number: SEK003 Storage Temperature: 2-8 C Fax : +86-10-58628220 Tel : +86-400-890-9989 http://www.sinobiological.com Description Mouse Monoclonal Antibody

More information

Strep-tag detection in Western blots

Strep-tag detection in Western blots Strep-tag detection in Western blots General protocol for the detection of Strep-tag fusion proteins Last date of revision April 2012 Version PR07-0010 www.strep-tag.com For research use only Important

More information

The Polymerase Chain Reaction. Chapter 6: Background

The Polymerase Chain Reaction. Chapter 6: Background The Polymerase Chain Reaction Chapter 6: Background PCR Amplify= Polymerase Chain Reaction (PCR) Invented in 1984 Applications Invention of PCR Kary Mullis Mile marker 46.58 in April of 1983 Pulled off

More information

SensoLyte Anti-alpha-Synuclein Quantitative ELISA Kit (Human/Mouse/Rat) *Colorimetric*

SensoLyte Anti-alpha-Synuclein Quantitative ELISA Kit (Human/Mouse/Rat) *Colorimetric* Catalog # Kit Size SensoLyte Anti-alpha-Synuclein Quantitative ELISA Kit (Human/Mouse/Rat) *Colorimetric* AS-55550 One 96-well strip plate This kit is optimized to detect human/mouse/rat alpha-synuclein

More information

Mouse TNF Alpha PicoKine ELISA Kit

Mouse TNF Alpha PicoKine ELISA Kit BOSTER BIOLOGICAL TECHNOLOGY 3942 B Valley Ave, Pleasanton, CA 94566 Phone: 888-466-3604 Fax: 925-215-2184 Email: boster@bosterbio.com Web: www.bosterbio.com Mouse TNF Alpha PicoKine ELISA Kit Catalog

More information

Updated PRODUCT INSERT. HPI Catalog # HP5-XXX

Updated PRODUCT INSERT. HPI Catalog # HP5-XXX Updated 2017 1 PRODUCT INSERT Hypoxyprobe, Inc. 121 Middlesex Turnpike Burlington, MA 01803 USA www.hypoxyprobe.com Hypoxyprobe Gemini Kit HPI Catalog # HP5-XXX Kit Contents: Solid pimonidazole HCl (Hypoxyprobe

More information

EAE Induction by Active Immunization in C57BL/6 Mice

EAE Induction by Active Immunization in C57BL/6 Mice Recommended protocol for use with: Hooke Kit MOG 35-55/CFA Emulsion PTX (EK-0110, EK-0112, EK-0113, EK- 0114, or EK-0115), or, Hooke Kit MOG 1-125/CFA Emulsion PTX (EK-0162 or EK-0164) Summary C57BL/6

More information

Production of Antisera to Synthetic Decapeptide of the Cooh-Terminus of Rat Glut2

Production of Antisera to Synthetic Decapeptide of the Cooh-Terminus of Rat Glut2 Production of Antisera to Synthetic Decapeptide of the Cooh-Terminus of Rat Glut2 Sandro Vestri 1, Hideki Nikami 2, Masayuki Saito 2 and Ubiratan F. Machado 1* 1 Department of Physiology and Biophysics,

More information

Anti-Piscirickettsia salmonis monoclonal antibody. Product no: P05

Anti-Piscirickettsia salmonis monoclonal antibody. Product no: P05 Anti-Piscirickettsia salmonis monoclonal antibody Product no: P05 Product Description The monoclonal antibody (Mab) against Piscirickettsia salmonis is specific for this bacterium. The specificity of the

More information

Human IL-1 Alpha PicoKine ELISA Kit

Human IL-1 Alpha PicoKine ELISA Kit BOSTER BIOLOGICAL TECHNOLOGY 3942 B Valley Ave, Pleasanton, CA 94566 Phone: 888-466-3604 Fax: 925-215-2184 Email: boster@bosterbio.com Web: www.bosterbio.com Human IL-1 Alpha PicoKine ELISA Kit Catalog

More information

E.Z.N.A. MicroElute Genomic DNA Kit. D preps D preps D preps

E.Z.N.A. MicroElute Genomic DNA Kit. D preps D preps D preps E.Z.N.A. MicroElute Genomic DNA Kit D3096-00 5 preps D3096-01 50 preps D3096-02 200 preps December 2013 E.Z.N.A. MicroElute Genomic DNA Kit Table of Contents Introduction...2 Kit Contents/Storage and Stability...3

More information

Therapeutic passive vaccination against chronic Lyme disease in mice

Therapeutic passive vaccination against chronic Lyme disease in mice Proc. Natl. Acad. Sci. USA Vol. 94, pp. 12533 12538, November 1997 Immunology Therapeutic passive vaccination against chronic Lyme disease in mice WEIMIN ZHONG*, THOMAS STEHLE*, CRISAN MUSETEANU*, ANNETTE

More information

IMMUNOPRECIPITATION (IP)

IMMUNOPRECIPITATION (IP) 1 IMMUNOPRECIPITATION (IP) Overview and Technical Tips 2 CONTENTS 3 7 8 9 12 13 17 18 19 20 Introduction Factors Influencing IP General Protocol Modifications Of IP Protocols Troubleshooting Contact Us

More information

GSI Equine IL-10 ELISA Kit- Cell Lysate DataSheet

GSI Equine IL-10 ELISA Kit- Cell Lysate DataSheet IL-10, also known as human cytokine synthesis inhibitory factor (CSIF), is an antiinflammatory cytokine that is produced by T cells, NK cells, mast cells and macrophages (1,2,3). It is capable of inhibiting

More information

Complement C4 Universal Kit

Complement C4 Universal Kit Order references Reagents CONT C4TUR-B00 Universal kit 1 x 50 ml R1 + 1 x 5 ml R2 C4TUR-00 Universal kit 2 x 50 ml R1 + 1 x 15 ml R2 C4TUR-C00 Universal kit 1 x 12 ml R1 + 1 x 2 ml R2 Other necessary products

More information

Product Information. Before you begin. Component A 1 vial of 30 ul vial of 300 ul each Glycerol. Tris

Product Information. Before you begin. Component A 1 vial of 30 ul vial of 300 ul each Glycerol. Tris Glowing Products for Science Mix-n-Stain Antibody Labeling Kits Size: 1 labeling per kit Storage: -20 o C Stability: Stable for at least 1 year from date of receipt when stored as recommended. Components:

More information

General Comments. Misinformation in Immunohistochemistry. Common Misinformation s in Immunohistochemistry 4/13/2017

General Comments. Misinformation in Immunohistochemistry. Common Misinformation s in Immunohistochemistry 4/13/2017 Common Misinformation s in Immunohistochemistry Tri State Meeting May 3, 2017 Steven Westra Reagent Product Specialist Leica Biosystems Misinformation in Immunohistochemistry Flood of New Markers Diagnostic

More information

ab Hypoxic Response Human Flow Cytometry Kit

ab Hypoxic Response Human Flow Cytometry Kit ab126585 Hypoxic Response Human Flow Cytometry Kit Instructions for Use For measuring protein levels by flow cytometry: hypoxia-inducible factor 1-alpha (HIF1A) and BCL2/adenovirus E1B 19 kda proteininteracting

More information

The Biotechnology Education Company. Quantitative ELISA. Storage: See Page 3 for specific storage instructions EXPERIMENT OBJECTIVE:

The Biotechnology Education Company. Quantitative ELISA. Storage: See Page 3 for specific storage instructions EXPERIMENT OBJECTIVE: The Biotechnology Education Company Revised and Updated Quantitative ELISA Storage: See Page 3 for specific storage instructions EXPERIMENT OBJECTIVE: EDVO-Kit # 278 The objective of this experiment is

More information

Electrophoresis and transfer

Electrophoresis and transfer Electrophoresis and transfer Electrophoresis Cation = positively charged ion, it moves toward the cathode (-) Anion = negatively charged ion, it moves toward the anode (+) Amphoteric substance = can have

More information

colorimetric sandwich ELISA kit datasheet

colorimetric sandwich ELISA kit datasheet colorimetric sandwich ELISA kit datasheet For the quantitative detection of mouse Interleukin 6 (IL6) concentrations in serum, plasma and cell culture supernatants. general information Catalogue Number

More information

One-Step Western TM Kit using TMB

One-Step Western TM Kit using TMB Technical Manual No. 0203 Version 03272008 I Description.. 1 II Kit Contents.. 2 III Applications 3 IV Key Features.. 3 V Storage.. 3 VI One-Step Western TM Protocol. 3 VII Examples. 3 VIII Troubleshooting..

More information

Virginia Western Community College BIO 205 General Microbiology

Virginia Western Community College BIO 205 General Microbiology Prerequisites BIO 205 General Microbiology One year of college biology and one year of college chemistry or divisional approval; an ENG 111 placement recommendation, co-enrollment in ENF 3/ENG 111, or

More information

Phagocytosis Assay Kit (IgG FITC)

Phagocytosis Assay Kit (IgG FITC) Phagocytosis Assay Kit (IgG FITC) Item No. 500290 Customer Service 800.364.9897 * Technical Support 888.526.5351 www.caymanchem.com TABLE OF CONTENTS GENERAL INFORMATION 3 Materials Supplied 4 Precautions

More information

Polyclonal ARHGAP25 antibody was prepared from rabbit serum after intracutaneous

Polyclonal ARHGAP25 antibody was prepared from rabbit serum after intracutaneous Preparation and purification of polyclonal antibodies Polyclonal ARHGAP25 antibody was prepared from rabbit serum after intracutaneous injections of glutathione S-transferase-ARHGAP25-(509-619) (GST-coiled

More information

A Role for Streptococcal Collagen-Like Protein 1 (Scl-1) in M1T1 Group A

A Role for Streptococcal Collagen-Like Protein 1 (Scl-1) in M1T1 Group A Supplemental Information for: A Role for Streptococcal Collagen-Like Protein 1 (Scl-1) in M1T1 Group A Streptococcus Resistance to Neutrophil Extracellular Traps Simon Döhrmann 1, Sabina Anik 1, Joshua

More information

PRODUCT INFORMATION. Composition of SOC medium supplied :

PRODUCT INFORMATION. Composition of SOC medium supplied : Product Name : Competent Cell BL21(DE3)pLysS Code No. : DS260 Size : 100 μl 10 Competency : > 5 10 7 cfu/μg (puc19) Supplied product : SOC medium, 1 ml 10 This product is for research use only Description

More information

SKIN INFECTION OF RABBITS WITH HEMOLYTIC STREP- TOCOCCI ISOLATED FROM A PATIENT WITH ERYSIPELAS.

SKIN INFECTION OF RABBITS WITH HEMOLYTIC STREP- TOCOCCI ISOLATED FROM A PATIENT WITH ERYSIPELAS. SKIN INFECTION OF RABBITS WITH HEMOLYTIC STREP- TOCOCCI ISOLATED FROM A PATIENT WITH ERYSIPELAS. I. METHOD OF DEMONSTRATING PROTECTIVE ACTION OF IMMUNE SERA. BY THOMAS M. RIVERS, M.D. (From the Hospital

More information

BrdU IHC Kit. For the detection and localization of bromodeoxyuridine incorporated into newly synthesized DNA of actively proliferating cells

BrdU IHC Kit. For the detection and localization of bromodeoxyuridine incorporated into newly synthesized DNA of actively proliferating cells K-ASSAY BrdU IHC Kit For the detection and localization of bromodeoxyuridine incorporated into newly synthesized DNA of actively proliferating cells Cat. No. KT-077 For Research Use Only. Not for Use in

More information

Sensitivity vs Specificity

Sensitivity vs Specificity Viral Detection Animal Inoculation Culturing the Virus Definitive Length of time Serology Detecting antibodies to the infectious agent Detecting Viral Proteins Western Blot ELISA Detecting the Viral Genome

More information

ab Ran Activation Assay Kit

ab Ran Activation Assay Kit ab173247 Ran Activation Assay Kit Instructions for Use For the simple and fast measurement of Ran activation. This product is for research use only and is not intended for diagnostic use. Version 1 Last

More information

Your Antibody Source. prosci-inc.com. Extensive Antibody Services Broad Antibody Catalog

Your Antibody Source. prosci-inc.com. Extensive Antibody Services Broad Antibody Catalog Your Source prosci-inc.com Extensive Services Broad Catalog Company Overview Established in 1998, ProSci Incorporated is a leading provider of high performance antibodies and custom antibody services.

More information

Mouse Galectin-3/ LGALS3 ELISA Kit

Mouse Galectin-3/ LGALS3 ELISA Kit Mouse Galectin-3/ LGALS3 ELISA Kit CATALOG NO: IRKTAH1395 LOT NO: SAMPLE INTENDED USE For quantitative detection of mouse Galectin-3 in cell culture supernates, serum and plasma (heparin, EDTA). BACKGROUND

More information

GUIDANCE ON THE EVALUATION OF NON ACCREDITED QUALIFICATIONS

GUIDANCE ON THE EVALUATION OF NON ACCREDITED QUALIFICATIONS GUIDANCE ON THE EVALUATION OF NON ACCREDITED QUALIFICATIONS 1. Introduction 1.1 This document provides guidance notes for the assessment of academic qualifications that have not been formally accredited

More information

Immunohistochemistry: Basics and Methods

Immunohistochemistry: Basics and Methods Immunohistochemistry: Basics and Methods Bearbeitet von Igor B Buchwalow, Werner Böcker 1st Edition. 2010. Buch. x, 153 S. Hardcover ISBN 978 3 642 04608 7 Format (B x L): 15,5 x 23,5 cm Gewicht: 445 g

More information

Supplemental Data Supplementary Figure Legends and Scheme Figure S1.

Supplemental Data Supplementary Figure Legends and Scheme Figure S1. Supplemental Data Supplementary Figure Legends and Scheme Figure S1. UTK1 inhibits the second EGF-induced wave of lamellipodia formation in TT cells. A and B, EGF-induced lamellipodia formation in TT cells,

More information

CytoGLOW. IKK-α/β. Colorimetric Cell-Based ELISA Kit. Catalog #: CB5358

CytoGLOW. IKK-α/β. Colorimetric Cell-Based ELISA Kit. Catalog #: CB5358 CytoGLOW IKK-α/β Colorimetric Cell-Based ELISA Kit Catalog #: CB5358 Please read the provided manual entirely prior to use as suggested experimental protocols may have changed. Research Purposes Only.

More information

FDA Perspective on the Preclinical Development of Cancer Vaccines

FDA Perspective on the Preclinical Development of Cancer Vaccines FDA Perspective on the Preclinical Development of Cancer Vaccines Richard D. McFarland Ph.D., M.D. Medical Officer CBER/OCTGT/DCEPT mcfarlandr@cber.fda.gov Cancer Vaccine Clinical Trials Workshop Alexandria,

More information

EGFR (Phospho-Ser695)

EGFR (Phospho-Ser695) Assay Biotechnology Company www.assaybiotech.com Tel: 1-877-883-7988 Fax: 1-877-610-9758 EGFR (Phospho-Ser695) Colorimetric Cell-Based ELISA Kit Catalog #: OKAG02090 Please read the provided manual entirely

More information

Western-GUARANTEED Antibody Service FAQ

Western-GUARANTEED Antibody Service FAQ Western-GUARANTEED Antibody Service FAQ Content Q 1: When do I need a Western GUARANTEED Peptide Antibody Package?...2 Q 2: Can GenScript provide a Western blot guaranteed antibody?...2 Q 3: Does GenScript

More information

Infection of Mice with Lyme Disease Spirochetes Constitutively Producing Outer Surface Proteins A and B

Infection of Mice with Lyme Disease Spirochetes Constitutively Producing Outer Surface Proteins A and B INFECTION AND IMMUNITY, June 2007, p. 2786 2794 Vol. 75, No. 6 0019-9567/07/$08.00 0 doi:10.1128/iai.01307-06 Copyright 2007, American Society for Microbiology. All Rights Reserved. Infection of Mice with

More information

INOS. Colorimetric Cell-Based ELISA Kit. Catalog #: OKAG00807

INOS. Colorimetric Cell-Based ELISA Kit. Catalog #: OKAG00807 INOS Colorimetric Cell-Based ELISA Kit Catalog #: OKAG00807 Please read the provided manual entirely prior to use as suggested experimental protocols may have changed. Research Purposes Only. Not Intended

More information

ab Mouse and Rabbit AP/Fast-Red (ABC) Detection IHC Kit

ab Mouse and Rabbit AP/Fast-Red (ABC) Detection IHC Kit ab128967 - Mouse and Rabbit AP/Fast-Red (ABC) Detection IHC Kit Instructions for Use For the detection of a specific antibody bound to an antigen in tissue sections. This product is for research use only

More information

Product Datasheet. TGN38 Antibody NBP Unit Size: 0.5 mg

Product Datasheet. TGN38 Antibody NBP Unit Size: 0.5 mg Product Datasheet TGN38 Antibody NBP1-03495 Unit Size: 0.5 mg Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles. Publications: 5 Protocols, Publications, Related Products,

More information

Assays for Immunogenicity: Are We There Yet?

Assays for Immunogenicity: Are We There Yet? Assays for Immunogenicity: Are We There Yet? Mark Wener, MD Department of Laboratory Medicine & Rheumatology Division Department of Medicine University of Washington Seattle, WA 98195 wener@uw.edu Goals:

More information

Mouse Factor XII Total ELISA Kit

Mouse Factor XII Total ELISA Kit Mouse Factor XII Total ELISA Kit Catalog No: IMFXIIKT-TOT Lot No: SAMPLE INTENDED USE This mouse coagulation Factor XII antigen assay is intended for the quantitative determination of total Factor XII

More information

Cloning and Sequence Analysis of the 22-kDa Antigen Genes of Orientia tsutsugamushi Strains Kato, TA763, AFSC 7, , TH1814, and MAK 119

Cloning and Sequence Analysis of the 22-kDa Antigen Genes of Orientia tsutsugamushi Strains Kato, TA763, AFSC 7, , TH1814, and MAK 119 Cloning and Sequence Analysis of the 22-kDa Antigen Genes of Orientia tsutsugamushi Strains Kato, TA763, AFSC 7, 18-032460, TH1814, and MAK 119 HONG GE, a MIN TONG, a ANDREW LI, b RAJAN MEHTA, b AND WEI-MEI

More information

Dr: RAWIA BADR Associate Professor of Microbiology&Immunology

Dr: RAWIA BADR Associate Professor of Microbiology&Immunology Dr: RAWIA BADR Associate Professor of Microbiology&Immunology Cell culture Commonly refers to the culture of animal cells and tissues, while the more specific term plant tissue.culture is used only for

More information

KPL SignaLOCK ChemiWestern Kits (Film and Imager Analysis)

KPL SignaLOCK ChemiWestern Kits (Film and Imager Analysis) KPL SignaLOCK ChemiWestern Kits (Film and Imager Analysis) SignaLOCK HRP ChemiWestern Kit (Film) Catalog No. 54-53-00 SignaLOCK HRP ChemiWestern Kit (Imager) Catalog No. 54-54-00 SignaLOCK AP ChemiWestern

More information

ab Mouse and Rabbit Specific HRP/DAB (ABC) Detection IHC Kit

ab Mouse and Rabbit Specific HRP/DAB (ABC) Detection IHC Kit ab64264 - Mouse and Rabbit Specific HRP/DAB (ABC) Detection IHC Kit Instructions for Use For the detection of a specific antibody bound to an antigen in tissue sections. This product is for research use

More information

mcherry Monoclonal Antibody (16D7) Catalog Number M11217 Product data sheet

mcherry Monoclonal Antibody (16D7) Catalog Number M11217 Product data sheet Website: thermofisher.com Customer Service (US): 1 800 955 6288 ext. 1 Technical Support (US): 1 800 955 6288 ext. 441 mcherry Monoclonal Antibody (16D7) Catalog Number M11217 Product data sheet Details

More information

Development of NOG mice

Development of NOG mice Development of NOG mice General characteris5cs of NOG mice 1. T and B cell deficient 2. NK cell deficient 3. Reduced macrophage and dendri;c cell func;on 4. Complement ac;vity deficient 5. No incidence

More information

Porcine IL-12/IL-23 p40 ELISA kit

Porcine IL-12/IL-23 p40 ELISA kit Porcine IL-12/IL-23 p40 ELISA kit Catalog number: NB-E50014 (96 wells) The kit is designed to quantitatively detect the levels of Porcine IL-12/IL-23 p40 in cell culture supernatants. FOR RESEARCH USE

More information