Colloids and Surfaces B: Biointerfaces

Save this PDF as:
 WORD  PNG  TXT  JPG

Size: px
Start display at page:

Download "Colloids and Surfaces B: Biointerfaces"

Transcription

1 Colloids and Surfaces B: Biointerfaces 97 (2012) Contents lists available at SciVerse ScienceDirect Colloids and Surfaces B: Biointerfaces jou rn al h om epage: Synthesis and cellular uptake of scatteredly cyclic RGDfK-conjugated superparamagnetic iron oxide nanoparticles Myung-Ik Yoo a,b, Yeong-Ju Seo c, Kyu-Sil Choi c, Jeong Sook Ha b, Kyoungja Woo a, a Molecular Recognition Research Center, Korea Institute of Science and Technology, P.O. Box 131, Cheongryang, Seoul , Republic of Korea b Department of Chemical and Biological Engineering, Korea University, Seoul , Republic of Korea c Molecular & Cellular Imaging Center, Samsung Biomedical Research Institute, Seoul , Republic of Korea a r t i c l e i n f o Article history: Received 7 October 2011 Received in revised form 22 March 2012 Accepted 5 April 2012 Available online 13 April 2012 Keywords: Superparamagnetic iron oxide nanoparticles (SPIONs) Scatteredly crgdfk-conjugated Ultrasmall Water-dispersible Lipophilic Cellular uptake a b s t r a c t We prepared scatteredly cyclic RGDfK-conjugated water-dispersible superparamagnetic iron oxide nanoparticles (crgdfk WSPIONs) and investigated their cellular uptake to MS-1 cells (mouse endothelial cell lines, express integrin v 3 ) and MCF-7 cells (human breast cancer cells, express low level of integrin v 3 ) using in vitro MRI. The crgdfk WSPIONs were prepared from oleate-protected SPIONs (SPION OA) as follows. Some oleates (OAs) on the SPION OA were substituted by mercaptohexadecanoic acids (MHA) and crgdfks were conjugated to MHAs. Finally, the remaining OAs were substituted by mercaptopropionic acids without detaching preexisting crgdfk-conjugated MHA ligands from the SPION surface. The crgdfk WSPIONs showed drastically higher cellular uptake than its corresponding control WSPIONs to MS-1 cells and also, showed higher selectivity to MS-1 cells than to MCF-7 cells, both implicating integrin v 3 -mediated cellular uptake of scatteredly crgdfk-conjugated WSPIONs Elsevier B.V. All rights reserved. 1. Introduction Surface engineering of superparamagnetic iron oxide nanoparticles (SPIONs) have attracted great attention since the SPIONs can be applied as a highly sensitive magnetic resonance imaging (MRI) agent utilizing T 2 contrast enhancement [1 7]. Commercial MRI agents using dextran-coated SPION clusters over 50 nm in hydrodynamic diameter (HD) are easily uptaken by the reticuloendothelial system (RES) and thus, have been used as a liver contrast agent. The clustered or aggregated nanoparticles greatly enhance their delivery to liver and spleen rather than the other target due to their much larger effective hydrodynamic size than the aggregation-free one [8,9]. Therefore, targeting and biocompatible SPIONs with HD below 10 nm in water are getting more important and challenging for noninvasive detection of tumors other than RES system at an early stage [7,10,11]. The core size below 10 nm is also required to ensure a superparamagnetism of the iron oxide nanoparticles without hysteresis, since the hysteresis induces magnetic agglomeration [2,12,13]. However, surface engineering of SPIONs (core size <10 nm) for targeting, biocompatible, aggregation-free, and water-soluble property has been a tough task, mostly because of Corresponding author. Tel.: , fax: address: (K. Woo). the multi-chelating hydrogen bonding between the nanoparticles during conjugation reaction in an aqueous solution [14 17]. It was a quite progress that Xie et al. [7] reported the targeting water-dispersible ultrasmall ( 8.4 nm) iron oxide nanoparticles based on the synthesis using 4-methylcatechol as a new surfactant and subsequent coupling of its aromatic ring to crgdyk via a Mannich reaction. Noteworthy is Tromsdorf et al. s [11] report on water-dispersible ultrasmall (HD < 10 nm) iron oxide nanoparticles based on PEGylation with phosphate functionalized PEG, though their materials do not include targeting molecules. Recently, starting from decanoate-protected Fe-rich SPIONs, we reported [18] the synthesis of scatteredly folate-conjugated lipophilic SPIONs (F LSPIONs) through scattered substitution of decanoate ligands for mercaptoundecanoic acid (MUA) and subsequent conjugation to folate. Here, we also reported [18] their folate receptor-mediated cellular uptake to KB cells (human epidermoid carcinoma cells, folate receptor positive) on a film made of F LSPIONs. This kind of targeting lipophilic SPIONs (LSPIONs) can be a nice candidate for T 2 contrast agent once they can be converted to aggregation-free water-dispersible ones. Meanwhile, we also reported [17] the preparation of scatteredly folate-conjugated quantum dot (QD) with HD below 10 nm by conversion of scatteredly folate-conjugated lipophilic QD to water-dispersible one. This water-dispersible conversion was possible through substitution of remaining decylamine ligands for mercaptopropionic acids /$ see front matter 2012 Elsevier B.V. All rights reserved.

2 176 M.-I. Yoo et al. / Colloids and Surfaces B: Biointerfaces 97 (2012) (MPAs) without detaching preexisting folate-conjugated MUA ligands from the QD surface. It was suggested that the targeting aggregation-free water-dispersible QDs in a physiological condition were obtainable by interrupting multi-chelating hydrogen bonding interactions between MPAs by using steric hindrance from the protruding targeting molecules [17]. This previous result encouraged us to expand the surface-engineering strategy of QDs to SPIONs and investigate their cellular interactions. In this report, PEG-bonded cyclic RGDfK (PEG crgdfk) was selected instead of folate as a targeting molecule to utilize its wellknown selectivity to integrin v 3, which is rich in the fibrinous cells and tumor tissues undergoing angiogenesis [19,20]. The PEG unit, which has been already bonded to crgdfk before conjugation to SPION, was employed for a biocompatibility [21,22]. However, its symbol PEG is frequently omitted in the abbreviated expression just for simplicity throughout this report. Our surface-engineering strategy worked nicely here with Fe-rich SPIONs like the case of Cd-rich QDs. The strategy is to substitute some oleates (OAs) of OA-protected Fe-rich SPION for mercaptohexadecanoic acids (MHAs) and subsequently conjugate crgdfk to MHA. Finally, the strategy converts scatteredly crgdfk-conjugated lipophilic SPIONs (crgdfk LSPIONs) to water-dispersible ones (crgdfk WSPIONs) by substitution of the remaining OAs for MPAs without detaching preexisting crgdfk-conjugated MHAs from the SPION surface. The crgdfk WSPIONs showed HD below 10 nm and integrin v 3 -mediated cellular uptake though the SPION was scatteredly crgdfk-conjugated. 2. Materials and methods 2.1. Synthesis of SPION( MPA) ex ( MHA apeg) 10 (WSPION) and SPION( MPA) ex ( MHA enpeg crgdfk) 10 (crgdfk WSPION) (material IV in Scheme 1) The PEG-conjugated and PEG crgdfk-conjugated LSPIONs, SPION( OA) ex ( MHA apeg) 10 and SPION( OA) ex ( MHA enpeg crgdfk) 10 (material III in Scheme 1, here ex is unknown excess amount [23]), were prepared according to the previous method [18] with some modifications using O- (2-aminoethyl)-O -methyl polyethylene glycol (apeg, M w = 2 kda, purchased from Fluka) and enpeg crgdfk (PEG M w = 600 Da, vide infra) and then, treated with MPA to transfer to aggregation-free WSPION and crgdfk WSPION. Briefly, Fe-rich SPIONs (8.5 nm by TEM image) protected with OA (SPION OA, I in Scheme 1) were prepared according to the published method [12,24] and saved as-prepared (about 22 ml) under Ar gas. A 3 ml portion of as-prepared Fe-rich SPION OA was washed with ethanol and acetonitrile consecutively, dispersed in toluene ( M, 6 ml), and kept as a stock under Ar atmosphere to preserve Fe-rich surface until it is used. A 2 ml portion of the stock solution was diluted to 30 ml and treated as follows. (1) MHA ( mmol in 0.15 ml toluene, roughly 10 eq. for SPION) was added to SPION OA ( mmol in 30 ml toluene) at 100 C and the solution was gently refluxed for 1 h under inert atmosphere. (2) To this solution at room temperature, dicyclohexylcarbodiimide (DCC, mmol in 0.15 ml toluene) was added and stirred for 1 h. Then, apeg or enpeg crgdfk ( mmol in 0.5 ml dimethylsulfoxide) was added and stirred overnight with intermittent treatments within 1 s in a sonication bath, under inert atmosphere and dark conditions. (3) The solution was transferred to a conical tube and a mixture (0.05 M, ml, 150 eq. for SPION) of MPA and NaOH in methanol was added and vortexed for 2 h with intermittent treatments within 1 s in a sonication bath. The short sonication within 1 s was helpful in steps (2) and (3) since it disturbs temporary weak agglomeration of nanoparticles Scheme 1. Surface modification process for aggregation-free, biocompatible, and targeting WSPION [I: SPION OA, II: SPION( OA) ex( MHA) 10, III: SPION( OA) ex( MHA enpeg crgdfk) 10 (crgdfk LSPION), IV: SPION( MPA) ex( MHA enpeg crgdfk) 10 (crgdfk-wspion)]. caused by poor solubility of conjugated crgdfk moiety and MPA in toluene. Here, 150 eq. of MPA was roughly chosen by considering the SPION surface area and the attachable MPA molecules, referring to the attached MPA molecules (80 100) to QD [25]. The resulting precipitate from step (3) was collected by centrifugation, rinsed with ethanol and then water, and finally dispersed in a ph 7.4 PBS solution to make WSPION and crgdfk WSPION solutions (20 ml each). The SPION samples were analyzed by FT-IR spectroscopy (Mattson IR 300) and TEM (Philips CM 30, 200 kv). The samples for FT-IR spectra were prepared as a pellet with KBr after fully dried. The HD was obtained from the Korea Basic Science Institute (Jeonju Center) using diluted samples ( M in PBS) with a particle size analyzer (UPA-150). The magnetic hysteresis curve of SPIONs was obtained from the Korea Basic Science Institute (Busan Center) using vibrating sample method (Princeton Measurement Co., Model 2900 Micromag) by measuring the precisely weighed sample in a folded parafilm (5 mm 5 mm) at room temperature. Fe concentrations of WSPION and crgdfk WSPION solutions were obtained after pretreatment using aqua regia from the Advanced Analysis Center of Korea Institute of Science and Technology using atomic absorption spectrophotometer (Thermo Electron Corp.) as 160 and 190 ppm, respectively. The biocompatible and integrin v 3 -targeting molecule, enpeg crgdfk, was prepared by coupling polyethylene glycol (PEG) derivative with crgdfk and ethylenediamine (en) as follows. First, PEG diacid ( mmol, M w = 600, Fluka) was dissolved in dried and distilled dimethylformamide (DMF, 10 ml). In an ice bath, the diacid groups were activated by slow addition of DCC (0.168 mmol in 1 ml DMF) and stirring for 16 h. Since the solution became translucent, additional DMF was added until the solid was dissolved (solution 1). Meanwhile, the crgdfk 2TFA (70 mg, mmol, M w = , 99.5%, purchased from FUTURECHEM Co. Ltd.) was dissolved in dried and distilled DMF (2 ml) and

3 M.-I. Yoo et al. / Colloids and Surfaces B: Biointerfaces 97 (2012) diisopropylamine (0.842 mmol, 99.95%, Aldrich) was added and stirred for 15 min to remove TFA from crgdfk. This solution was slowly added to solution 1 and stirred for 3 h to make an amide bond between crgdfk and one end of activated PEG diacid. Then, en was added and stirred for 1 h to make an additional amide bond between en and the other end of activated PEG diacid. Diethyl ether was added to collect enpeg crgdfk precipitate. The precipitate was rinsed with methylene chloride, dried with vacuum, filled with Ar gas, and saved in a freezer (87.5 mg, 90.6% yield). The amide bond formation at both ends of PEG with en and crgdfk was confirmed from the FT-IR spectrum (Supplementary data) In vitro characterization of vˇ3-specific binding to WSPION and crgdfk WSPION In order to examine whether the scatteredly crgdfk-conjugated WSPIONs specifically bind to integrin v 3, MS-1 cells (mouse endothelial cell line) which express v 3 and MCF-7 cells (human breast cancer cell line) which express relatively low level of v 3 were chosen. Expression of integrin v 3 in MS-1 and MCF-7 was examined by western blot analysis using anti- v and 3 antibodies purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA) and Cell Signaling Technology, Inc. (Danvers, MA), respectively. The expression of -actin as a control was also examined with its monoclonal antibody purchased from Santa Cruz Biotechnology, Inc (Santa Cruz, CA). The cells were cultured in DMEM supplemented with 10% fetal bovine serum until 90% confluence and then incubated with 20 g Fe/mL of WSPION and crgdfk WSPION for 2 h in the media. The cells were washed with PBS three times, scraped with rubber policeman and suspended in PBS ( cells/ml). An aliquot of cells was analyzed for the content of iron bound to cells by ICP-MS (Agilent 7500A, Palo Alto, CA) and the rest of cells were imbedded in 1% agarose for MR imaging (Philips Healthcare, Achieva, 3.0 T). T 2 weighted MR imaging was performed in TR/TE = 5000/100 ms. The binding experiment was also done with Prussian blue staining to visualize the content of iron. MS-1 and MCF-7 cells were subconfluently cultured on the chamber slide and 20 g Fe/mL of WSPION and crgdfk WSPION for 2 h in the media. Cells were then washed three times with PBS, treated with 4% paraformaldehyde solution at 4 C for 30 min to fix the cells, and washed three times with PBS again. A 1:1 mixture of 5% potassium ferrocyanide (II) trihydrate solution and 5% HCl was added to each well and the cells were incubated at room temperature for 1 h before being counterstained with nuclear fast red for 5 min. Each well was then washed three times with PBS and analyzed by light microscopy Cytotoxicity of crgdfk WSPION The in vitro cytotoxic effect of crgdfk WSPION was tested in HT1080 (Human fibrosarcoma), MCF-7 and MS-1 cell lines using a 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay kit (R&D Systems, Minneapolis, MN, USA). Briefly, subconfluent cells in a 96-well plate were incubated with various concentrations (0.1 1 M) of crgdfk WSPION at 37 C for 12 h. As a cytotoxic control, cell lysis buffer (RIPA buffer) was also added with serial dilution. An MTT reagent was added to the culture media, and the cells were left at 37 C for 3 h. The cells were treated with detergent solution and were incubated at 20 C for 4 h in the dark. The absorbance of the well was measured at 570 nm with a reference wavelength of 650 nm in a plate reader (Spectra Max M2; Molecular Devices, Sunnyvale, CA, USA). Chart 1. Molecular structure of apeg and enpeg crgdfk. 3. Results and discussion Scheme 1 illustrates the stepwise surface modification process for WSPIONs and crgdfk WSPIONs. (1) The initial SPION OA (I) with Fe-rich surface was treated with 10 eq. of MHA, replacing OA at appropriately scattered places through Fe S linkage. (2) The next step was to make a simple conjugation to incoming functional molecule, apeg for WSPIONs and enpeg crgdfk for crgdfk WSPIONs (see Chart 1 for the structure), through amide linkage. Here, we assumed that most of the anchored MHA ligands were conjugated to apeg or enpeg crgdfk since the conjugation is an easy working reaction and we used 3 eq. of activating agent and apeg or enpeg crgdfk per MHA molecule. (3) Finally, the SPIONs were treated with MPA to substitute OA remaining on the LSPION surface, yielding aggregation-free WSPIONs and crgdfk WSPIONs. In this report, we have chosen cyclic RGD peptide unit as a targeting molecule instead of folate unit of the previous lipophilic SPION [18], since cyclic RGD peptide is well known as a specific marker of integrin v 3 which is rich in fibrinous cells and tumor tissues undergoing angiogenesis [19,20]. In step (1), roughly 10 eq. of MHA per SPION ( 8.5 nm) was chosen as a compromising point considering inter-nanoparticle magnetic interaction and our previous report [17] for QDs ( 6 nm), which showed the aggregation-free upper limit 15 eq. of MUA. The double bond in the middle of OA backbone has been known to be saturated during heating for SPION OA synthesis [26]. Thiolate anchoring occurred nicely when the SPION surface was Fe-rich [18] whereas there seemed to be no anchoring when its surface was oxidized. The surface oxidation can be indirectly judged from the following conjugation step since there is no conjugation to incoming amine-functionalized molecules in case of the oxidized SPIONs due to the lack of mediating MHA molecule. In step (3), like the cases of QDs [17] and gold nanoparticles [27], the mercapto-functionalized short-chain hydrophilic molecules readily substituted OA ligands, whereas they did not substitute the mercapto-functionalized long-chain hydrocarbon ligands on the Fe-rich SPION surface because of their (MPA here) weaker coordinating power. It was indirectly evidenced through a ligand exchange test as shown in Supplementary data. Here, for a comparison, we have used dodecanthiol (DDT) instead of MHA since fully MHA-protected SPION MHA was seriously aggregated and not dispersible in any solvent due to multi-chelating hydrogen bonding. The TEM images in Figure S2 indicate replacement of OA by DDT from slightly closer distances between SPION DDTs ( 2.0 nm) in b than SPION OAs ( 2.7 nm) in a, however, no replacement of DDT

4 178 M.-I. Yoo et al. / Colloids and Surfaces B: Biointerfaces 97 (2012) Fig. 2. Hysteresis curve and photo image (inset) of crgdfk WSPION. Fig. 1. FT-IR spectra of SPIONs according to Scheme 1. (a) I: SPION OA, (b) II: SPION( OA) ex( MHA) 10, (c) III: SPION( OA) ex( MHA enpeg crgdfk) 10, (d) IV: SPION( MPA) ex( MHA enpeg crgdfk) 10, (e) enpeg crgdfk. by MPA from little change in inter-particle distances in c. The photo images in Figure S3 display the dispersion property of the SPIONs according to the ligand exchange test. The hydrophobic SPION OA and SPION DDT are dispersible in toluene whereas the hydrophilic SPION MPA should be dispersible in water if SPION DDT has been converted to SPION MPA. However, the SPION DDT has not transferred to water layer even after treatment with MPA. On the other hand, it has been known that Fe-rich SPION OA can be directly converted to SPION MPA by treatment with MPA [24]. Therefore, it is suggested that the mercapto-functionalized short-chain hydrophilic molecules readily substitute OA ligands whereas they cannot substitute the mercapto-functionalized long-chain hydrocarbon ligands on the Fe-rich SPION surface like the cases of QDs [17] and gold nanoparticles [27], since the longer chain provides the higher coordinating power to its terminal thiol group through higher electron density. Fig. 1 shows the FT-IR spectra of SPIONs according to the process of Scheme 1. Spectrum a for SPION OA (I) indicates the existence of iron oxide with oleate, which shows C H stretching bands just below 3000 cm 1, carbonyl band at 1702 cm 1, CH 2 and CH 3 wagging bands at 1465 cm 1 and 1376 cm 1, iron carboxylate bands around 1434 cm 1, C O stretching band at 1115 cm 1 and Fe O band around 603 cm 1 [24]. Spectrum b for scatteredly MHA substituted SPION OA (II) shows increased carboxylate bands around 1425 cm 1 and broad O H stretching band from H-bonded carboxylic acid around 3436 cm 1. The Fe O band at around 605 cm 1 has been relatively increased since more OA has been washed out during centrifugation for purification. Spectrum c displays the conjugation of enpeg crgdfk to SPION. The bands around 1026 cm 1 and 1658 cm 1 indicate C O stretching modes from PEG and amide C O bands from crgdfk, respectively. As a reference, the FT- IR spectrum of enpeg crgdfk is shown in e. Spectrum d shows Fig. 3. (a) TEM images of SPIONs according to Scheme 1 and (b) hydrodynamic diameter distribution of WSPION (A) and crgdfk WSPION (B).

5 M.-I. Yoo et al. / Colloids and Surfaces B: Biointerfaces 97 (2012) Fig. 4. (a) Western blot analysis, (b) T 2-weighed MR images, (c) relative intensity of T 2 image, (d) SPION uptake, (e) Prussian blue staining of MS-1 and MCF-7 cells after culture with no SPION ( ), WSPION (A) and crgdfk WSPION (B). *P < 0.05, n = 3 for (c) and (d). (For interpretation of the references to color in this figure legend, the reader is referred to the web version of the article.) decrease of C H stretching band just below 3000 cm 1 and increase of H-bonded carboxylic acid O H bands around 3440 cm 1 and H- bonded C O bands at 1644 cm 1, due to substitution of remaining OA by MPA. Fig. 2 shows the hysteresis curve of crgdfk WSPION, including a photo image of its solution as an inset. The coercivity and remanence values are not discernible at room temperature, showing superparamagnetic behavior. The saturation magnetization value 40 emu/g was slightly lower than that (42 emu/g) [12] for 11 nm maghemite nanoparticles with OA surfactants as expected from their size 8.5 nm. This lower value compared to the bulk maghemite (73.5 emu/g) [12] is likely due to the mass of organic molecules bonded to the SPION surface, whereas the bulk maghemite does not have any organic molecules. It implicates that the magnetic property has been preserved after all the surface engineering process. The crgdfk WSPION solution was a dark brown homogeneous solution and the crgdfk WSPION solid was collectable by an external magnet. Although it precipitated after long time, it recovered its original dispersible property by simple shaking.

6 180 M.-I. Yoo et al. / Colloids and Surfaces B: Biointerfaces 97 (2012) Fig. 3(a) shows the TEM images of each SPION according to the process of Scheme 1. The inter-particle distances in the TEM image I indicate the spaces occupied by OA on the SPION. When some of the OA was substituted by MHA, there could be weak H- bonding attractions between the particles through carboxylic acid groups. However, the SPIONs are still separated by the organic molecules in all the solutions up to III and IV, which correspond to enpeg crgdfk conjugated lipophilic and water-dispersible SPIONs, respectively. Fig. 3(b) shows the hydrodynamic diameter distribution of WSPION (A) and crgdfk WSPION (B) solution. They show quite narrow size distribution and the average hydrodynamic diameter of A and B were 9.9 and 9.4 nm, respectively, implicating that they are aggregation-free in aqueous solution. The zeta potential of WSPION was ± 0.92 mev and explains its waterdispersible property. MS-1 and MCF-7 cells were chosen for cellular uptake test since MS-1 cells highly express v 3 whereas MCF-7 cells express relatively low level of v 3. Fig. 4(a) shows western blot analysis using anti- v and 3 antibodies, and -actin as a control. Both cells showed relatively low level of v expression, although MCF-7 cells showed slightly higher v level than MS-1 cells. On the other hand, MS-1 cells showed high level of 3 expression whereas MCF-7 cells showed almost undetectable level of 3 expression. Fig. 4(b) shows the T 2 -weighed MR images of both cells cultured in the SPION-free (, control), A, and B solutions. As expected, the darkness of T 2 image contrast was B > A > control in both MS-1 and MCF-7 cells. However, MS-1 cells showed much lower T 2 intensity than MCF-7 cells as expected from the western blot analysis. The relative intensity of T 2 image is shown in Fig. 4(c). The relative intensity of T 2 image was analyzed in the software, Image J. The intensity of signal decrease was B > A > control in both MS-1 and MCF-7 cells and the difference of signal decrease was much higher in MS-1 cells than MCF-7 cells as expected. The SPION uptake by the cells was analyzed by the Fe content per cell and displayed in Fig. 4(d). The SPION uptake from B was more than twice of that from A in case of MS-1 cells, whereas it showed only slight difference in case of MCF-7 cells. The SPION uptake from B was much higher in MS-1 cells than MCF-7 cells. The Fe content of cells cultured without SPION was almost undetectable in both cells. The cells cultured in SPION-free, A, and B solutions were visualized by Prussian blue staining and shown in Fig. 4(e). The blue spots were rarely observed in the both cells cultured without SPION. In case of MS-1 cells, the prominent blue spots were observed when the cells were cultured with B whereas the blue spots were greatly reduced when cultured with A. In case of MCF-7 cells, there was only a slight difference in the blue spots with a slightly higher intensity in B than A. All these results confirm that the cellular uptake of B was mediated by integrin v 3 binding. The cytotoxic effect of crgdfk WSPION was investigated with MTT assay in HT1080 (human fibrosarcoma cells), MCF-7, and MS- 1 cells. The cell lysis buffer (RIPA) was used as a control of cell killing. The cytotoxic effect of crgdfk WSPION on the tested cells was minimal as shown in Fig. 5. The SPION did not show any significant cytotoxicity to the three kinds of cells in the tested range of concentration. In summary, we have prepared ultrasmall (HD < 10 nm) nontoxic aggregation-free water-dispersible scatteredly crgdfk (specific marker of integrin v 3 )-conjugated SPIONs starting from oleate-protected SPIONs. The preparation was done stepwise by substitution of some oleates of SPION OA for mercaptohexadecanoic acids and then, conjugation to crgdfk and finally, conversion to water-dispersible ones by substitution of remaining oleate surfactants for mercaptopropionic acids. The PEGconjugated SPIONs without crgdfk have been prepared for a control experiment. The crgdfk-conjugated SPIONs showed higher cellular uptake to MS-1 cells than MCF-7 cells and also, showed Fig. 5. MTT assay of HT1080 (, ), MS-1 (, ), and MCF-7 (, ) cells with RIPA (,, ) and crgdfk WSPION (,, ). drastically higher uptake to MS-1 cells than the control SPIONs, confirming integrin v 3 -mediated cellular uptake of the scatteredly crgdfk-conjugated SPIONs. Acknowledgment This research was supported by Future Key Technology Program of Korea Institute of Science and Technology and by Future-based Technology Development Program (Green Nano Technology Development Program) through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology (grant number ). Appendix A. Supplementary data Supplementary data associated with this article can be found, in the online version, at j.colsurfb References [1] Y. Zhang, N. Kohler, M. Zhang, Biomaterials 23 (2002) [2] J.H. Lee, Y.M. Huh, Y. Jun, J. Seo, J. Jang, H.T. Song, S. Kim, E.J. Cho, H.G. Yoon, J.S. Suh, J. Cheon, Nat. Med. 13 (2007) 95. [3] Y.W. Jun, Y.M. Huh, J.S. Choi, J.H. Lee, H.T. Song, S. Kim, S. Yoon, K.S. Kim, J.S. Shin, J.S. Suh, J. Cheon, J. Am. Chem. Soc. 127 (2005) [4] J. Qin, S. Laurent, Y.S. Jo, A. Roch, M. Mikhaylova, Z.M. Bhujwalla, R.N. Muller, M. Muhammed, Adv. Mater. 19 (2007) [5] S. Rasaneh, H. Rajabi, M.H. Babaei, S. Akhlaghpoor, J. Nanopart. Res. 13 (2011) [6] H.J. Chung, H. Lee, K.H. Bae, Y. Lee, J. Park, S.-W. Cho, J.Y. Hwang, H. Park, R. Langer, D. Anderson, T.G. Park, ACS Nano 5 (2011) [7] J. Xie, K. Chen, H.-Y. Lee, C. Xu, A.R. Hsu, S. Peng, X. Chen, S. Sun, J. Am. Chem. Soc. 130 (2008) [8] S.M. Moghimi, A.C. Hunter, J.C. Murray, Pharm. Rev. 53 (2001) 283. [9] T. Neuberger, B. Schopf, H. Hofmann, M. Hofmann, B. von Rechenberg, J. Magn. Magn. Mater. 293 (2005) 483. [10] H.S. Choi, W. Liu, P. Misra, E. Tanaka, J.P. Zimmer, B.I. Ipe, M.G. Bawendi, J.V. Frangioni, Nat. Biotechnol. 25 (2007) [11] U.I. Tromsdorf, O.T. Bruns, S.C. Salmen, U. Beisiegel, H. Weller, Nano Lett. 9 (2009) [12] K. Woo, J. Hong, S. Choi, H.-W. Lee, J.-P. Ahn, C.S. Kim, S.W. Lee, Chem. Mater. 16 (2004) [13] E. Schellenberger, J. Schnorr, C. Reutelingsperger, L. Ungethum, W. Meyer, M. Taupitz, B. Hamm, Small 4 (2008) 225. [14] J. Ge, Y. Hu, M. Biasini, W.P. Beyermann, Y. Yin, Angew. Chem. Int. Ed. 46 (2007) [15] H. Ai, C. Flask, B. Weinverg, X.-T. Shuai, M.D. Pagel, D. Farrell, J. Duerk, J. Gao, Adv. Mater. 17 (2005) [16] H.-B. Xia, J. Yi, P.-S. Foo, B. Liu, Chem. Mater. 19 (2007) [17] J. Moon, K.-S. Choi, B. Kim, K.-H. Yoon, T.-Y. Seong, K. Woo, J. Phys. Chem. C 113 (2009) 7114.

7 M.-I. Yoo et al. / Colloids and Surfaces B: Biointerfaces 97 (2012) [18] K. Woo, J. Moon, K.-S. Choi, T.-Y. Seong, K.-H. Yoon, J. Magn. Magn. Mater. 321 (2009) [19] Z.-F. Su, G. Liu, S. Gupta, Z. Zhu, M. Rusckowski, D. Hnatowich, Bioconjug. Chem. 13 (2002) 561. [20] Z.B. Li, W.B. Cai, Q.Z. Cao, K. Chen, Z.H. Wu, L.N. He, X.Y. Chen, J. Nucl. Med. 48 (2007) [21] M.D. Bentley, M.J. Bossard, K.W. Burton, T.S. Viegas, Mod. Biopharm. 4 (2005) [22] A.M. Derfus, W.C. Chan, S.N. Bhatia, Adv. Mater. 16 (2004) 961. [23] Since the number of OA molecules on SPION-OA is quite variable according to the washing condition, it is not certain how many OAs are remained after ten MHAs replace ten OAs on SPION I. Knowing that there are large excess amount of OA molecules per SPION-OA by thermal analysis, we assumed that there are still large excess amount of OAs on the SPION II in Scheme 1. So, ten MHA-anchored and then, enpeg crgdfk-conjugated or apeg-conjugated lipophilic SPION III was designated respectively as SPION( OA)ex( MHA enpeg crgdfk)10 and SPION( OA)ex( MHA apeg)10. [24] Y.T. Lee, K. Woo, K.-S. Choi, IEEE Trans. Nanotechnol. 7 (2008) 111. [25] W. Cai, D.-W. Shin, K. Chen, O. Gheysens, Q. Cao, S.X. Wang, S.S. Gambhir, X. Shen, Nano Lett. 6 (2006) 669. [26] A.L. Willis, N.J. Turro, S. O Brien, Chem. Mater. 17 (2005) [27] R.S. Ingram, M.J. Hostetler, R.W. Murray, J. Am. Chem. Soc. 119 (1997) 9175.

ACS Nano, Article ASAP DOI: /nn800632j

ACS Nano, Article ASAP DOI: /nn800632j Synthesis, Characterization, and Bioconjugation of Fluorescent Gold Nanoclusters toward Biological Labeling Applications Walter H. Chang et. al. Department of Biomedical Engineering, Chung Yuan Christian

More information

nanoprecipitation mpeg-pla 2 nd Emulsion mpeg-pla in DCM

nanoprecipitation mpeg-pla 2 nd Emulsion mpeg-pla in DCM THERAPEUTIC NANOTECHNOLOGY LAB MODULE Location: BioNano Lab, 3119 Micro and Nanotechnology Laboratory (MNTL) Instructor: Jianjun Cheng, Assistant Professor of Materials Science and Engineering Lab Assistants:

More information

Fe doped Magnetic Nanodiamonds made by Ion

Fe doped Magnetic Nanodiamonds made by Ion Fe doped Magnetic Nanodiamonds made by Ion Implantation ChienHsu Chen a, I.C. Cho b, Hui-Shan Jian c and H. Niu a* a Nuclear Science and Technology Development Center, National Tsing Hua University, HsinChu

More information

Supporting Information

Supporting Information Electronic Supplementary Material (ESI) for Materials Chemistry Frontiers. This journal is the Partner Organisations 2017 Supporting Information Supramolecular Conjugated Polymer Materials for Organelle

More information

A Robust Procedure for the Functionalization of Gold Nanorods and Noble Metal Nanoparticles

A Robust Procedure for the Functionalization of Gold Nanorods and Noble Metal Nanoparticles A Robust Procedure for the Functionalization of Gold Nanorods and Noble Metal Nanoparticles Benjamin Thierry,*, Jane Ng, Tina Krieg, and Hans J. Griesser Ian Wark Research Institute, University of South

More information

ph-responsive gold nanoparticles-in-liposome hybrid nanostructures for

ph-responsive gold nanoparticles-in-liposome hybrid nanostructures for Electronic Supplementary Information ph-responsive gold nanoparticles-in-liposome hybrid nanostructures for enhanced systemic tumor delivery Jutaek Nam, a Yeong Su Ha, b Sekyu Hwang, a Woonghee Lee, b

More information

A New Theranostic System Based on Gold Nanocages and Phase-Change Materials with Unique Features for Photoacoustic Imaging and Controlled Release

A New Theranostic System Based on Gold Nanocages and Phase-Change Materials with Unique Features for Photoacoustic Imaging and Controlled Release 1/2011 Supporting Information A New Theranostic System Based on Gold Nanocages and Phase-Change Materials with Unique Features for Photoacoustic Imaging and Controlled Release Geon Dae Moon,, Sung-Wook

More information

Compact Zwitterion-Coated Iron Oxide Nanoparticles for Biological Applications

Compact Zwitterion-Coated Iron Oxide Nanoparticles for Biological Applications Compact Zwitterion-Coated Iron Oxide Nanoparticles for Biological Applications The MIT Faculty has made this article openly available. Please share how this access benefits you. Your story matters. Citation

More information

AnaTag HiLyte Fluor 647 Protein Labeling Kit

AnaTag HiLyte Fluor 647 Protein Labeling Kit AnaTag HiLyte Fluor 647 Protein Labeling Kit Catalog # 72049 Kit Size 3 Conjugation Reactions This kit is optimized to conjugate HiLyte Fluor 647 SE to proteins (e.g., IgG). It provides ample materials

More information

tel: IRON OXIDE-BASED SUPERPARAMAGNETIC CONTRAST AGENTS

tel: IRON OXIDE-BASED SUPERPARAMAGNETIC CONTRAST AGENTS tel:4006551678 email: fige007@163.com IRON OXIDE-BASED SUPERPARAMAGNETIC CONTRAST AGENTS Molday ION TM product line is a family of iron oxide-based superparamagnetic contrast reagents designed to label

More information

Supporting Information for. Nitric Oxide Reactivity of Copper(II) Complexes of Bidentate Amine Ligands: Effect of. Substitution on Ligand Nitrosation

Supporting Information for. Nitric Oxide Reactivity of Copper(II) Complexes of Bidentate Amine Ligands: Effect of. Substitution on Ligand Nitrosation 1 Supporting Information for Nitric Oxide Reactivity of Copper(II) Complexes of Bidentate Amine Ligands: Effect of Substitution on Ligand Nitrosation Moushumi Sarma and Biplab Mondal Department of Chemistry,

More information

TACS MTT Assays. Cell Proliferation and Viability Assays. Catalog Number: TA tests. Catalog Number: TA tests

TACS MTT Assays. Cell Proliferation and Viability Assays. Catalog Number: TA tests. Catalog Number: TA tests TACS MTT Assays Cell Proliferation and Viability Assays Catalog Number: TA5355-2500 tests Catalog Number: TA5412-5000 tests This package insert must be read in its entirety before using this product. FOR

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

for New Energy Materials and Devices; Beijing National Laboratory for Condense Matter Physics,

for New Energy Materials and Devices; Beijing National Laboratory for Condense Matter Physics, Electronic Supplementary Information Highly efficient core shell CuInS 2 /Mn doped CdS quantum dots sensitized solar cells Jianheng Luo, a Huiyun Wei, a Qingli Huang, a Xing Hu, a Haofei Zhao, b Richeng

More information

The One Year Fate of Iron Oxide Coated Gold. Nanoparticles in Mice

The One Year Fate of Iron Oxide Coated Gold. Nanoparticles in Mice Supporting Information The One Year Fate of Iron Oxide Coated Gold Nanoparticles in Mice Jelena Kolosnjaj-Tabi, 1,2+ Yasir Javed, 3+ Lénaic Lartigue, 1,3 Jeanne Volatron, 1 Dan Elgrabli, 1 Iris Marangon,

More information

Enzyme-mediated preparation of hydrogels composed of poly(ethylene glycol) and gelatin as cell culture platforms

Enzyme-mediated preparation of hydrogels composed of poly(ethylene glycol) and gelatin as cell culture platforms Electronic Supplementary Material (ESI) for RSC Advances. This journal is The Royal Society of Chemistry 2014 Supplementary Material (ESI) for RSC Advances Enzyme-mediated preparation of hydrogels composed

More information

Supplementary information for. An Ultrasensitive Biosensor for DNA Detection Based on. Hybridization Chain Reaction Coupled with the Efficient

Supplementary information for. An Ultrasensitive Biosensor for DNA Detection Based on. Hybridization Chain Reaction Coupled with the Efficient Supplementary information for An Ultrasensitive Biosensor for DNA Detection Based on Hybridization Chain Reaction Coupled with the Efficient Quenching of Ruthenium Complex to CdTe Quantum Dot Yufei Liu,

More information

MitoBiogenesis In-Cell ELISA Kit (Colorimetric)

MitoBiogenesis In-Cell ELISA Kit (Colorimetric) PROTOCOL MitoBiogenesis In-Cell ELISA Kit (Colorimetric) DESCRIPTION 1850 Millrace Drive, Suite 3A Eugene, Oregon 97403 MS643 Rev.2 For identifying inhibitors and activators of mitochondrial biogenesis

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

Total Histone H3 Acetylation Detection Fast Kit (Fluorometric)

Total Histone H3 Acetylation Detection Fast Kit (Fluorometric) Total Histone H3 Acetylation Detection Fast Kit (Fluorometric) Catalog Number KA1539 48 assays Version: 02 Intended for research use only www.abnova.com Table of Contents Introduction... 3 Intended Use...

More information

Engineering Quantum Dots for Live-Cell Single-Molecule Imaging

Engineering Quantum Dots for Live-Cell Single-Molecule Imaging Engineering Quantum Dots for Live-Cell Single-Molecule Imaging Andrew M. Smith and Shuming Nie Georgia Tech and Emory University Department of Biomedical Engineering 2011 NSF Nanoscale Science and Engineering

More information

This journal is The Royal Society of Chemistry S 1

This journal is The Royal Society of Chemistry S 1 2013 S 1 Thermochemical analysis on the growth of NiAl 2 O 4 rods Sang Sub Kim, a Yong Jung Kwon, b Gunju Sun, a Hyoun Woo Kim,* b and Ping Wu* c a Department of Materials Science and Engineering, Inha

More information

RayBio Human, Mouse and Rat Phospho-STAT3 (Tyr705) ELISA Kit

RayBio Human, Mouse and Rat Phospho-STAT3 (Tyr705) ELISA Kit RayBio Human, Mouse and Rat Phospho-STAT3 (Tyr705) ELISA Kit Catalog #: PEL-Stat3-Y705 User Manual Last revised August 10, 2016 Caution: Extraordinarily useful information enclosed ISO 13485 Certified

More information

Polyvidone Polyvinylpyrrolidone H 2 C H C N

Polyvidone Polyvinylpyrrolidone H 2 C H C N 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 (C 6 H 9 NO)n [9003-39-8] Poly [(2-oxo-1-pyrrolidinyl) ethylene] Povidone (Rev. 1, Stage 4)

More information

TECHNICAL BULLETIN. Catalog Number RAB0012 Storage Temperature 20 C

TECHNICAL BULLETIN. Catalog Number RAB0012 Storage Temperature 20 C Phospho-Akt (pser 473 )/pan-akt ELISA Kit for detection of human, mouse, or rat phospho-akt (pser 473 ) and pan-akt in cell and tissue lysates Catalog Number RAB0012 Storage Temperature 20 C TECHNICAL

More information

For the development of sandwich ELISAs to measure phosphorylated Vascular Endothelial Growth Factor Receptor 1 (VEGF R1/Flt-1) in cell lysates.

For the development of sandwich ELISAs to measure phosphorylated Vascular Endothelial Growth Factor Receptor 1 (VEGF R1/Flt-1) in cell lysates. DuoSet IC Human Phospho-VEGF R1/Flt-1 Catalog Number DYC4170-2 Catalog Number DYC4170-5 For the development of sandwich ELISAs to measure phosphorylated Vascular Endothelial Growth Factor Receptor 1 (VEGF

More information

Colorimetric detection of influenza A (H1N1) virus based on peptide functionalized polydiacetylene (PEP-PDA) nanosensor

Colorimetric detection of influenza A (H1N1) virus based on peptide functionalized polydiacetylene (PEP-PDA) nanosensor Electronic Supplementary Material (ESI) for RSC Advances. This journal is The Royal Society of Chemistry 2016 Electronic Supporting Information Colorimetric detection of influenza A (H1N1) virus based

More information

Supporting information

Supporting information Electronic Supplementary Material (ESI) for ChemComm. This journal is The Royal Society of Chemistry 2015 Supporting information Near Infrared Light-responsive and Injectable Supramolecular Hydrogels for

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

Whole Mount IHC Protocol

Whole Mount IHC Protocol Whole Mount IHC Protocol Authors: Ruth Sullivan, Ryan Trevena and Kyle Wegner Creation Date: 03/17/2016 All steps should be conducted with gentle agitation on an orbital shaker, unless otherwise instructed.

More information

RayBio Human, Mouse and Rat Phospho-STAT3 (Tyr705) and Total STAT3 ELISA Kit

RayBio Human, Mouse and Rat Phospho-STAT3 (Tyr705) and Total STAT3 ELISA Kit RayBio Human, Mouse and Rat Phospho-STAT3 (Tyr705) and Total STAT3 ELISA Kit Catalog #: PEL-Stat3-Y705-T User Manual Last revised October 10, 2017 Caution: Extraordinarily useful information enclosed ISO

More information

Correlation between the Structure and Catalytic. Activity of [Cp*Rh(Substituted Bipyridine)] Complexes for NADH Regeneration

Correlation between the Structure and Catalytic. Activity of [Cp*Rh(Substituted Bipyridine)] Complexes for NADH Regeneration Supporting Information Correlation between the Structure and Catalytic Activity of [Cp*Rh(Substituted Bipyridine)] Complexes for NADH Regeneration Vinothkumar Ganesan, Dharmalingam Sivanesan and Sungho

More information

RayBio Phospho- Stat 3 (Tyr705) ELISA Kit

RayBio Phospho- Stat 3 (Tyr705) ELISA Kit RayBio Phospho- Stat 3 (Tyr705) ELISA Kit For Measuring Phosphorylated Stat3 (Tyr705) in Human, Mouse and Rat Cell Lysates User Manual (Revised Mar 1, 2012) RayBio Stat3 (Tyr705) ELISA Kit Protocol (Cat#:

More information

ab BrdU Immunohistochemistry Kit

ab BrdU Immunohistochemistry Kit ab125306 - BrdU Immunohistochemistry Kit Instructions for Use For the detection and localization of bromodeoxyuridine incorporated into newly synthesized DNA of actively proliferating cells. This product

More information

64 CuCl 2 in 50 µl 0.1N NaOAc buffer, and 20 µg of each DOTA-antibody conjugate in 40 µl

64 CuCl 2 in 50 µl 0.1N NaOAc buffer, and 20 µg of each DOTA-antibody conjugate in 40 µl Number of DOTA per antibody The average number of DOTA chelators per antibody was measured using a reported procedure with modifications (1,2). Briefly, nonradioactive CuCl 2 (80-fold excess of DOTA antibodies)

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

Novel concept of rechargeable battery using iron oxide nanorods. anode and nickel hydroxide cathode in aqueous electrolyte

Novel concept of rechargeable battery using iron oxide nanorods. anode and nickel hydroxide cathode in aqueous electrolyte Supplementary Information for: Novel concept of rechargeable battery using iron oxide nanorods anode and nickel hydroxide cathode in aqueous electrolyte Zhaolin Liu *, Siok Wei Tay and Xu Li Institute

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

For identifying inhibitors and activators of mitochondrial biogenesis in adherent cultured cells.

For identifying inhibitors and activators of mitochondrial biogenesis in adherent cultured cells. ab110216 MitoBiogenesis TM In-Cell ELISA Kit (IR) Instructions for Use For identifying inhibitors and activators of mitochondrial biogenesis in adherent cultured cells. This product is for research use

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

RayBio Phospho- Akt (Ser473) ELISA Kit

RayBio Phospho- Akt (Ser473) ELISA Kit RayBio Phospho- Akt (Ser473) ELISA Kit For Measuring Phosphorylated Akt (Ser473) in Human, Mouse and Rat Cell Lysates User Manual (Revised Mar 1, 2012) RayBio Akt (Ser473) ELISA Kit Protocol (Cat#: PEL-Akt-S473-001)

More information

Ultrathin Nanosheets of Feroxyhyte: A New Two-dimensional. Hefei National Laboratory for Physical Sciences at Microscale,

Ultrathin Nanosheets of Feroxyhyte: A New Two-dimensional. Hefei National Laboratory for Physical Sciences at Microscale, Electronic Supplementary Material (ESI) for Chemical Science. This journal is The Royal Society of Chemistry 2014 Electronic Supplementary Information for Ultrathin Nanosheets of Feroxyhyte: A New Two-dimensional

More information

Mouse Luteinizing Hormone (LH) ELISA

Mouse Luteinizing Hormone (LH) ELISA Mouse Luteinizing Hormone (LH) ELISA For the quantitative determination of mouse LH in serum, plasma and tissue homogenates Cat. No. KU-222 For Research Use Only. Not for use in diagnostic procedures.

More information

ab GFP ELISA Kit Instructions for Use For the quantitative measurement of GFP protein expression

ab GFP ELISA Kit Instructions for Use  For the quantitative measurement of GFP protein expression ab117992 GFP ELISA Kit Instructions for Use For the quantitative measurement of GFP protein expression This product is for research use only and is not for diagnostic use. intended www.abcam.com Table

More information

In vitro Human Umbilical Vein Endothelial Cells (HUVEC) Tube-formation Assay. Josephine MY Ko and Maria Li Lung *

In vitro Human Umbilical Vein Endothelial Cells (HUVEC) Tube-formation Assay. Josephine MY Ko and Maria Li Lung * In vitro Human Umbilical Vein Endothelial Cells (HUVEC) Tube-formation Assay Josephine MY Ko and Maria Li Lung * Clinical Oncology Department, The Univerisity of Hong Kong, Hong Kong, Hong Kong SAR *For

More information

ProductInformation. Genomic DNA Isolation Kit. Product No. GDI-3 Technical Bulletin No. MB-275 May 2000 TECHNICAL BULLETIN

ProductInformation. Genomic DNA Isolation Kit. Product No. GDI-3 Technical Bulletin No. MB-275 May 2000 TECHNICAL BULLETIN Genomic DNA Isolation Kit Product No. GDI-3 Technical Bulletin No. MB-275 May 2000 TECHNICAL BULLETIN ProductInformation Product Description Sigma s Genomic DNA Isolation Kit isolates genomic DNA from

More information

SERVA Ni-NTA Magnetic Beads

SERVA Ni-NTA Magnetic Beads INSTRUCTION MANUAL SERVA Ni-NTA Magnetic Beads Magnetic beads for Affinity Purification of His-Tag Fusion Proteins (Cat. No. 42179) SERVA Electrophoresis GmbH - Carl-Benz-Str. 7-69115 Heidelberg Phone

More information

This Document Contains:

This Document Contains: This Document Contains: 1. In-Cell Western Protocol II. Cell Seeding and Stimulation Supplemental Protocol III. Complete Assay Example: Detailing the Seeding, Stimulation and Detection of the A431 Cellular

More information

ab Histone H3 Acetylation Assay Kit

ab Histone H3 Acetylation Assay Kit ab115102 Histone H3 Acetylation Assay Kit Instructions for Use For the measurement of global histone H3 acetylation using a variety of mammalian cells This product is for research use only and is not intended

More information

Supporting Information

Supporting Information Supporting Information A Highly Selective Mitochondria-Targeting Fluorescent K + Sensor Xiangxing Kong, Fengyu Su, Liqiang Zhang, Jordan Yaron, Fred Lee, Zhengwei Shi, Yanqing Tian,* and Deirdre R. Meldrum*

More information

METHCATHINONE Latest Revision: July 11, 2005

METHCATHINONE Latest Revision: July 11, 2005 METHCATHINONE Latest Revision: July 11, 2005 O CH 3 HN CH 3 1. SYNONYMS CFR: Methcathinone CAS #: Base: 5650-44-2 Hydrochloride: 49656-78-2 Other Names: N-methylcathinone Methylcathinone 2-Methylamino-1-phenyl-1-propanone

More information

StemXVivo. Mesoderm Kit. Catalog Number SC030B. Reagents for the differentiation of human pluripotent stem cells into mesoderm.

StemXVivo. Mesoderm Kit. Catalog Number SC030B. Reagents for the differentiation of human pluripotent stem cells into mesoderm. StemXVivo Mesoderm Kit Catalog Number SC030B Reagents for the differentiation of human pluripotent stem cells into mesoderm. This package insert must be read in its entirety before using this product.

More information

ab Complex IV Human Enzyme Activity Microplate Assay Kit

ab Complex IV Human Enzyme Activity Microplate Assay Kit ab109909 Complex IV Human Enzyme Activity Microplate Assay Kit Instructions for Use For the quantitative measurement of Complex IV activity in samples from Human and Cow. This product is for research use

More information

Human Pluripotent Stem Cell Functional Identification Kit

Human Pluripotent Stem Cell Functional Identification Kit Human Pluripotent Stem Cell Functional Identification Kit Catalog Number SC027B Reagents for the identification of human pluripotent stem cells by in vitro functional differentiation. This package insert

More information

E.Z.N.A. Tissue DNA Kit. D preps D preps D preps

E.Z.N.A. Tissue DNA Kit. D preps D preps D preps E.Z.N.A. Tissue DNA Kit D3396-00 5 preps D3396-01 50 preps D3396-02 200 preps August 2016 E.Z.N.A. Tissue DNA Kit Table of Contents Introduction and Overview...2 Yield and Quality of DNA...3 Illustrated

More information

DNA isolation from tissue DNA isolation from eukaryotic cells (max. 5 x 106 cells) DNA isolation from paraffin embedded tissue

DNA isolation from tissue DNA isolation from eukaryotic cells (max. 5 x 106 cells) DNA isolation from paraffin embedded tissue INDEX KIT COMPONENTS 3 STORAGE AND STABILITY 3 BINDING CAPACITY 3 INTRODUCTION 3 IMPORTANT NOTES 4 EUROGOLD TISSUE DNA MINI KIT PROTOCOLS 5 A. DNA isolation from tissue 5 B. DNA isolation from eukaryotic

More information

Preparation of cerium oxide nanoparticles (CNPs). Preparations of CNPs produced

Preparation of cerium oxide nanoparticles (CNPs). Preparations of CNPs produced Electronic Supplemental Information Preparation of cerium oxide nanoparticles (CNPs). Preparations of CNPs produced from two synthetic procedures were tested that have been previously described 11. CNPs

More information

Porcine Transferrin Receptor(TFR) ELISA Kit

Porcine Transferrin Receptor(TFR) ELISA Kit Porcine Transferrin Receptor(TFR) ELISA Kit Catalog No. CSB-E13481p (96T) This immunoassay kit allows for the in vitro quantitative determination of porcine TFR concentrations in serum, plasma. Expiration

More information

Swelling Behavior Study of γ-irradiated Gelatin Hydrogels Prepared in Organic/Aqueous Mixtures

Swelling Behavior Study of γ-irradiated Gelatin Hydrogels Prepared in Organic/Aqueous Mixtures J. Ind. Eng. Chem., Vol. 13, No. 6, (2007) 997-1001 Swelling Behavior Study of γ-irradiated Gelatin Hydrogels Prepared in Organic/Aqueous Mixtures Junhwa Shin, Youn Mook Lim, Joon-Pyo Jeun, and Young Chang

More information

PROTOCOL. Monoamine Oxidase B Enzyme Specific Activity Assay Kit (human) MS747 Rev.0 DESCRIPTION ADDITIONAL MATERIALS REQUIRED

PROTOCOL. Monoamine Oxidase B Enzyme Specific Activity Assay Kit (human) MS747 Rev.0 DESCRIPTION ADDITIONAL MATERIALS REQUIRED PROTOCOL Monoamine Oxidase B Enzyme Specific Activity Assay Kit (human) 1850 Millrace Drive, Suite 3A Eugene, Oregon 97403 MS747 Rev.0 DESCRIPTION MAOB Enzyme Specific Activity Assay Kit Sufficient materials

More information

In situ generation of Li 2 FeSiO 4 coating on MWNT as a high rate cathode material for lithium ion batteries

In situ generation of Li 2 FeSiO 4 coating on MWNT as a high rate cathode material for lithium ion batteries Supporting Information: In situ generation of Li 2 FeSiO 4 coating on MWNT as a high rate cathode material for lithium ion batteries Yi Zhao, Jiaxin Li, Ning Wang, Chuxin Wu, Yunhai Ding, Lunhui Guan*

More information

ab Histone H4 Acetylation Assay Kit

ab Histone H4 Acetylation Assay Kit ab115103 Histone H4 Acetylation Assay Kit Instructions for Use For the measurement of global histone H4 acetylation using a variety of mammalian cells, including Tissue, adherent and suspension cells This

More information

UV-Induced DNA Damage ELISA (CPD Quantitation)

UV-Induced DNA Damage ELISA (CPD Quantitation) KAMIYA BIOMEDICAL COMPANY UV-Induced DNA Damage ELISA (CPD Quantitation) For the rapid detection and quantitation of CPD in any DNA samples Cat. No. KT-914 For Research Use Only. Not for use in diagnostic

More information

Instructions. Fuse-It-siRNA. Shipping and Storage. Overview. Kit Contents. Specifications. Note: Important Guidelines

Instructions. Fuse-It-siRNA. Shipping and Storage. Overview. Kit Contents. Specifications. Note: Important Guidelines Membrane fusion is a highly efficient method for transfecting various molecules and particles into mammalian cells, even into sensitive and primary cells. The Fuse-It reagents are cargo-specific liposomal

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

Characterization of mab aggregation using a Cary 60 UV-Vis Spectrophotometer and the Agilent 1260 Infinity LC system

Characterization of mab aggregation using a Cary 60 UV-Vis Spectrophotometer and the Agilent 1260 Infinity LC system Characterization of mab aggregation using a Cary 60 UV-Vis Spectrophotometer and the Agilent 1260 Infinity LC system Application Note Biopharmaceuticals Authors Arunkumar Padmanaban and Sreelakshmy Menon

More information

MECHANICAL PROPERTIES OF SURFACTANT-COATING CARBON NANOFIBER/EPOXY COMPOSITE

MECHANICAL PROPERTIES OF SURFACTANT-COATING CARBON NANOFIBER/EPOXY COMPOSITE International Journal of Nanoscience, Vol. 1, Nos. 5 & 6 (2002) 1 6 c World Scientific Publishing Company MECHANICAL PROPERTIES OF SURFACTANT-COATING CARBON NANOFIBER/EPOXY COMPOSITE ZHE YING,, JIN-HONG

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

» Talc is a native, hydrous magnesium silicate, sometimes containing a small proportion of aluminum silicate.

» Talc is a native, hydrous magnesium silicate, sometimes containing a small proportion of aluminum silicate. Change to read: Talc» Talc is a native, hydrous magnesium silicate, sometimes containing a small proportion of aluminum silicate. Packaging and storage Preserve in well closed containers. Identification

More information

Electronic Supplementary Information

Electronic Supplementary Information Electronic Supplementary Information Ultrasensitive immunoassay based on dual signal amplification of the electrically heated carbon electrode and QDs functionalized labels for the detection of matrix

More information

MATERIAL DATA SHEET. Reagents Provided in Kit

MATERIAL DATA SHEET. Reagents Provided in Kit Lot # XXXXX MATERIAL DATA SHEET HSP70/HSP40 Glow-Fold Protein Refolding Kit Cat. # K-290 Heat shock proteins (HSPs) are a family of highly conserved stress response proteins. Heat shock proteins function

More information

MSD Immuno-Dot-Blot Assays. A division of Meso Scale Diagnostics, LLC.

MSD Immuno-Dot-Blot Assays. A division of Meso Scale Diagnostics, LLC. MSD Immuno-Dot-Blot Assays Example: High Throughput Western Blots Replacements Traditional Western Blots High content Molecular weight and immunoreactivity Labor and protein intensive Inherently low throughput

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

SensoLyte pnpp Alkaline Phosphatase Assay Kit *Colorimetric*

SensoLyte pnpp Alkaline Phosphatase Assay Kit *Colorimetric* SensoLyte pnpp Alkaline Phosphatase Assay Kit *Colorimetric* Revision Number: 1.1 Last updated: October 2014 Catalog # Kit Size AS-72146 500 Assays (96-well plate) Optimized Performance: This kit is optimized

More information

Cytotoxicity LDH Assay Kit-WST

Cytotoxicity LDH Assay Kit-WST Cytotoxicity LDH Assay Kit-WST Supplementary Information Notice to Users Preparation of Reagent This instruction complements the Technical Manual in the product. Please use this instruction as supplements

More information

Supplementary Material (ESI) for Chemical Communications This journal is (c) The Royal Society of Chemistry 2009

Supplementary Material (ESI) for Chemical Communications This journal is (c) The Royal Society of Chemistry 2009 Supplementary Information Silver Nanoparticles with Planar Twinned Defects: Effect of Halides for Precise Tuning of Plasmon Absorption from 400 to >900 nm by Nicole Cathcart, Andrew J. Frank and Vladimir

More information

ab CFSE Fluorescent Cell Labeling Kit

ab CFSE Fluorescent Cell Labeling Kit ab113853 CFSE Fluorescent Cell Labeling Kit Instructions for Use For the durable fluorescent labeling of live cells for fluorescent microscopy and flow cytometry, population growth studies and within sample

More information

DNA-RNA EXTRACTION. Dr. Amira A. T. AL-Hosary Lecturer of infectious diseases, Faculty of Veterinary Medicine, Assiut University, Egypt

DNA-RNA EXTRACTION. Dr. Amira A. T. AL-Hosary Lecturer of infectious diseases, Faculty of Veterinary Medicine, Assiut University, Egypt DNA-RNA EXTRACTION Dr. Amira A. T. AL-Hosary Lecturer of infectious diseases, Faculty of Veterinary Medicine, Assiut University, Egypt Nucleic Acids (DNA & RNA) DNA and RNA Breaks (Nucleotides) I. DNA

More information

HUMAN ENDOTHELIAL- CELL SPECIFIC MOLECULE- 1 (ESM-1) ELISA KIT

HUMAN ENDOTHELIAL- CELL SPECIFIC MOLECULE- 1 (ESM-1) ELISA KIT PAGE 1 HUMAN ENDOTHELIAL- CELL SPECIFIC MOLECULE- 1 (ESM-1) ELISA KIT FOR THE QUANTITATIVE DETERMINATION OF HUMAN ESM-1 CONCENTRATIONS IN SERUM AND EDTA PLASMA ALWAYS REFER TO LOT SPECIFIC PROTCOL PROVIDED

More information

EPIGENTEK. EpiQuik In Situ Histone H3-K27 Tri-Methylation Assay Kit. Base Catalog # P-3014T PLEASE READ THIS ENTIRE USER GUIDE BEFORE USE

EPIGENTEK. EpiQuik In Situ Histone H3-K27 Tri-Methylation Assay Kit. Base Catalog # P-3014T PLEASE READ THIS ENTIRE USER GUIDE BEFORE USE EpiQuik In Situ Histone H3-K27 Tri-Methylation Assay Kit Base Catalog # PLEASE READ THIS ENTIRE USER GUIDE BEFORE USE The EpiQuik In Situ Histone H3-K27 Tri-Methylation Assay Kit is suitable for specifically

More information

Supporting Information. Photoinduced Anion Exchange in Cesium Lead Halide Perovskite Nanocrystals

Supporting Information. Photoinduced Anion Exchange in Cesium Lead Halide Perovskite Nanocrystals Supporting Information Photoinduced Anion Exchange in Cesium Lead Halide Perovskite Nanocrystals David Parobek, Yitong Dong, Tian Qiao, Daniel Rossi, Dong Hee Son* Department of Chemistry, Texas A&M University,

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

ab MetaPath Mito Disease 4-Plex Dipstick Array

ab MetaPath Mito Disease 4-Plex Dipstick Array ab109879 MetaPath Mito Disease 4-Plex Dipstick Array Instructions for Use For the measurement of mitochondrial biogenesis regulation in human samples This product is for research use only and is not intended

More information

Electronic Supplementary Information (ESI) for. In vivo Two-photon Fluorescent Imaging of Fluoride with a Desilylationbased Reactive Probe

Electronic Supplementary Information (ESI) for. In vivo Two-photon Fluorescent Imaging of Fluoride with a Desilylationbased Reactive Probe Electronic Supplementary Information (ESI) for In vivo Two-photon Fluorescent Imaging of Fluoride with a Desilylationbased Reactive Probe Dokyoung Kim, a Subhankar Singha, a Taejun Wang, b Eunseok Seo,

More information

HOOK Sulfo-NHS-Biotin

HOOK Sulfo-NHS-Biotin 126PR-03 G-Biosciences 1-800-628-7730 1-314-991-6034 technical@gbiosciences.com A Geno Technology, Inc. (USA) brand name HOOK Sulfo-NHS-Biotin For the coupling of biotin to protein amine groups (Cat. #

More information

Q. sample preparation for FTIR & How to sample introduce in the system?

Q. sample preparation for FTIR & How to sample introduce in the system? Q. sample preparation for FTIR & How to sample introduce in the system? FTIR SAMPLE PREPARATION FTIR sample cell designed according to nature of a sample. Sample may be liquid/solid/gaseous. Sample preparation

More information

PARP-1 (cleaved) Human In-Cell ELISA Kit (IR)

PARP-1 (cleaved) Human In-Cell ELISA Kit (IR) ab110215 PARP-1 (cleaved) Human In-Cell ELISA Kit (IR) Instructions for Use For the quantitative measurement of Human PARP-1 (cleaved) concentrations in cultured adherent and suspension cells. This product

More information

Canine Symmetric dimethylarginine ELISA Kit

Canine Symmetric dimethylarginine ELISA Kit Distribuito in ITALIA da Li StarFish S.r.l. Via Cavour, 35 20063 Cernusco S/N (MI) telefono 02-92150794 fax 02-92157285 info@listarfish.it www.listarfish.it Optimize Your Research Canine Symmetric dimethylarginine

More information

ab G alpha i Activation Assay Kit

ab G alpha i Activation Assay Kit ab173234 G alpha i Activation Assay Kit Instructions for Use For the simple and fast measurement of G alpha i activation. This product is for research use only and is not intended for diagnostic use. Version

More information

Bovine Prostaglandin E2 (PG-E2) ELISA Kit

Bovine Prostaglandin E2 (PG-E2) ELISA Kit Bovine Prostaglandin E2 (PG-E2) ELISA Kit Catalog Number. CSB-E14237B For the quantitative determination of endogenic bovine prostaglandin E2 (PG-E2) concentrations in serum, plasma, tissue homogenates.

More information

Phospho-Progesterone Receptor (S294) Cell-Based

Phospho-Progesterone Receptor (S294) Cell-Based Phospho-Progesterone Receptor (S294) Cell-Based Colorimetric ELISA Kit Catalog No. KA1327C Detection and Quantification of Phospho-Progesterone Receptor (S294) Protein Concentration in Cell. Research Purposes

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

PROTOCOL 1850 Millrace Drive, Suite 3A Eugene, Oregon

PROTOCOL 1850 Millrace Drive, Suite 3A Eugene, Oregon PROTOCOL Complex I Enzyme Activity Dipstick Assay Kit 1850 Millrace Drive, Suite 3A Eugene, Oregon 97403 MS130 Rev.3 DESCRIPTION Complex I Enzyme Activity Dipstick Assay Kit Sufficient materials are provided

More information

Data Sheet. PD-1[Biotinylated]:PD-L2 Inhibitor Screening Assay Kit Catalog # Size: 96 reactions

Data Sheet. PD-1[Biotinylated]:PD-L2 Inhibitor Screening Assay Kit Catalog # Size: 96 reactions Data Sheet PD-1[Biotinylated]:PD-L2 Inhibitor Screening Assay Kit Catalog # Size: 96 reactions DESCRIPTION: Cell signaling through the PD-1 receptor upon binding the PD-L2 ligand attenuates immune responses

More information

SYNTHESIS OF MAGNETITE NANO-PARTICLES BY REVERSE CO- PRECIPITATION

SYNTHESIS OF MAGNETITE NANO-PARTICLES BY REVERSE CO- PRECIPITATION 2nd International Conference on Ultrafine Grained & Nanostructured Materials (UFGNSM) International Journal of Modern Physics: Conference Series Vol. 5 (2012) 160 167 World Scientific Publishing Company

More information

Protocol. VeriKine TM Human Interferon Alpha Multi-Subtype Serum ELISA Kit

Protocol. VeriKine TM Human Interferon Alpha Multi-Subtype Serum ELISA Kit Protocol VeriKine TM Human Interferon Alpha Multi-Subtype Serum ELISA Kit Catalog No: 41110 Assay Range: 12.5-1000 pg/ml Store all components at 2 8 C Sold under license from Pestka Biomedical Laboratories,

More information

Extraction of DNA staining dyes from DNA using hydrophobic ionic liquids

Extraction of DNA staining dyes from DNA using hydrophobic ionic liquids Electronic Supplementary Information Extraction of DNA staining dyes from DNA using hydrophobic ionic liquids Imran Khimji, Krystina Doan, Kiara Bruggeman, Po-Jung Jimmy Huang, Puja Vajha, and Juewen Liu*

More information

NAG-1 (GDF-15, MIC-1) Prostate Cancer ELISA kit

NAG-1 (GDF-15, MIC-1) Prostate Cancer ELISA kit NAG-1 (GDF-15, MIC-1) Prostate Cancer ELISA kit Catalog Number: NG1 Store at -0 C. FOR RESEARCH USE ONLY v. 1081 Introduction This sandwich ELISA kit is for determination of NAG-1 (GDF-15, MIC-1) levels

More information

Synthesis of High Magnetic Moment CoFe Nanoparticles via Interfacial Diffusion in Core/Shell Structured Co/Fe Nanoparticles

Synthesis of High Magnetic Moment CoFe Nanoparticles via Interfacial Diffusion in Core/Shell Structured Co/Fe Nanoparticles Nano Res (2009) 2: 380 3859 DOI 10.1007/s12274-009-9037-4 Research Article 00380 Synthesis of High Magnetic Moment CoFe Nanoparticles via Interfacial Diffusion in Core/Shell Structured Co/Fe Nanoparticles

More information

LDH-Cytox Assay Kit. A Colorimetric Cytotoxicity Measuring Kit. Cat. No LDH-Cytox Assay Kit can be used to measure cytotoxicity in vitro

LDH-Cytox Assay Kit. A Colorimetric Cytotoxicity Measuring Kit. Cat. No LDH-Cytox Assay Kit can be used to measure cytotoxicity in vitro A Colorimetric Cytotoxicity Measuring Kit Cat. No. 426401 LDH-Cytox Assay Kit can be used to measure cytotoxicity in vitro BioLegend, Inc Biolegend.com It is highly recommended that this manual be read

More information