Nordic. MUbio. FIX&PERM Cell Fixation and Permeabilization Kit. Find more at nordicmubio.com

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1 FIX&PERM Cell Fixation and Permeabilization Kit FIX&PERM is a simple procedure making use of two reagents. Reagent A gently fixes cells, while Reagent B permeabilizes them. Reagent A gently fixes cells preserving their forward and side-scatter characteristics Reagent B and labelled antibodies are then added The specific formulation of FIX&PERM reduces background while allowing the simultaneous addition of permeabilization medium and fluorochrome labelled antibodies for staining of membrane proteins as well as intracellular components, such as cytoplasmic enzymes, nuclear proteins, oncoproteins, cytokines, immunoglobulins, and so on. Mildly ixes cells, preserving their low cytometric scatter characteristics Allows simultaneous characterisation of both intracellular and cell surface markers Rapid technique whole procedure can be carried out in less than one hour, ready for immediate analysis or storage for 24 hours Reagent B permeabilizes the cell Stringent QC procedures the quality of each lot is determined using well-defined blood samples and subsequent comparison of immunostaining and scatter characteristics of obtained leukocyte populations, ensuring consistent and reliable results lot after lot A range of antibodies to intracellular and membrane antigens with optimised protocols for use with FIX&PERM Allowing binding and detection of intracellular antigens Visit now to find the products you need for your immunological research, or get in touch on info@nordicmubio.com

2 Procedure For each sample to be analysed use 50 µl of whole blood, bone marrow or mononuclear cell suspension in a 5 ml tube Add 100 µl of Reagent A (Fixation Medium, stored and used at room temperature) Incubate for 15 minutes at room temperature Add 5 ml phosphate buffered saline and centrifuge cells for 5 minutes at 300 g Remove supernatant and add to cell pellet 100 µl Reagent B and 20 µl of the appropriate monoclonal antibody FIX&PERM Cell Fixation and Permeabilization Kit Size Cat. Number Fix&PERM Kit 2 x 20 ml GAS-002 Fix&PERM 1000 Kit 2 x 100 ml GAS Fix&PERM Reagent A (Fix) 1 x 100 ml GAS-002A-1 Fix&PERM Reagent B (Perm) 1 x 100 ml GAS-002B-1 Fix&PERM Sample Kit GAS-002M 2 x 5 ml All FIX&PERM kits are CE Marked The majority of available monoclonal antibody conjugates can be used with FIX&PERM. However, in some cases the fixation step may require optimization. - offers a number of mouse anti-human conjugated (PE/FITC) monoclonal antibodies pre-optimised for use with FIX&PERM. Antigen Isotype Size Cat. Number Vortex at low speed for 1-2 seconds CD3 IgG1 Incubate for 15 minutes at room temperature CD22 IgG1 MPO-C2 IgG1 0.2 mg 0.2 mg 0.2 mg GM-4011 GM-4012 (FITC) GM-4013 (PE) GM-4051 GM-4052 (FITC) GM-4053 (PE) GM-4191 GM-4192 (FITC) GM-4193 (PE) Wash cells with PBS as above Remove supernatant and resuspend cells in sheath luid for immediate analysis Alternativly, resuspend cells in 0.5 ml 1.0 % formaldehyde and store at 2-8 C in the dark. Analyse cells within 24 hours Mouse anti-human Combi Products IgG Negative Control (FITC) and IgG Negative Control (PE) Anti-Lysozyme (FITC) and anti-lactoferrin (PE) GIC-201 GIC-206 Anti-Myeloperoxidase-C2 (FITC) and anti-lactoferrin (PE) GIC-212 Anti-Myeloperoxidase-C2 (FITC) and anti-cd3 (PE) GIC-213 Anti-Myeloperoxidase-C2 (FITC) and anti-cd22 (PE) GIC-214 More products with protocols optimised for FIX&PERM can be found here: Contact your local distributor here:

3 High Quality Green Fluorescent Protein (GFP) Antibodies Green Fluorescent Protein (GFP) is a 27kD protein derived from the Aequorea victoria (A. victoria) jellyish. It is a naturally luorescent protein which has revolutionised life science and has been used to study proteins in a wide range of applications. GFP has been used extensively to monitor gene expression; to localise proteins at the cellular and sub-cellular level; to track metastatic cells and as a tag to identify proteins in vitro. - is pleased to announce the release of a new range of products directed against the native form of GFP. Our new range of anti-gfp antibodies bring a number of advantages over antibodies produced against recombinant, or variant, forms of GFP. This includes the ability to detect GFP produced from a wide range of diferent sources, including native, recombinant, and a number of diferent mutants, without sufering from high levels of non-speciic binding/background. Part of the reason many anti-recombinant GFP antibodies sufer from high background issues is due to contamination of an innoculum with other E.coli components, leading to increased non-speciic binding. Our products were tested for cross-reactivity to a wide range of antigens, including many derived from E.coli, and demonstrated no cross-reactivity. A. victoria is a bioluminescent jellyish, or hydromedusa, found of the west coast of North America. The enormous interest in GFP as a protein marker has given rise to the development of polyclonal and monoclonal antibodies against GFP, used for the immunolocalisation and immunoblotting of GFPfused proteins in a variety of applications. Since its discovery, many variants of GFP have been engineered making antibodies recognising a wide range of these variants an important, and useful, scientiic tool for use alongside GFP-fused proteins. Catalogue Number Bii-nGFPab2-100 Bii-nGFPab3-100 Bii-nGFPab4-100 Bii-nGFPab4-PO Bii-nGFPabC1-100 Bii-rGFP-100 Description Rabbit anti-gfp (native) Rabbit anti-gfp (native) Rabbit anti-gfp (native) Rabbit anti-gfp (native):po Chicken anti-gfp (native) Recombinant Green Fluorescent Protein (wild type) Format Ammonium Sulfate fraction Protein A Puriied Ainity Puriied Soyabean Peroxidase Conjugate Ammonium Sulfate fraction Recombinant Protein Unit Size 0.1 ml

4 How do we know our GFP products are the highest quality? Example Size Exclusion Chromatography (HPLC) Results Bii-nGFPab3-100 (Protein A puriied) Competitor D To ensure - s new GFP antibody range is of the highest quality a number of comparison experiments were run with the bestselling antibodies from other suppliers against recombinant GFP, assessing purity, speciicity, and titre of the antibodies. This page presents a number of the indings from these anlayses. Bii-nGFPab3-100 (Protein A puriied) Competitor E The HPLC analyses of the - ainity puriied antibodies show just a single peak, illustrating their purity. The competitor antibodies with multiple peaks have contaminants, such as serum albumin, which can afect the results of e.g. Western blots or ELISAs. Competitor A Competitor B - Native GFP antibody Anti-GFP Ab source Native GFP Antibody HPLC Purity SDS Purity Western Speciicity Clean Clean Clean Competitor A Contaminants Contaminants Competitor B Clean Clean Competitor C Competitor D Multiple peaks Many peaks (contains BSA) Western blot Anlaysis The gels in these images have been loaded with E.coli control, not expressing GFP (lane A), E.coli expressing GFP (lane B), and molecular weight standards (lane C). These images show intact GFP (main band at 27 kda), with some high and low mol. weight bands related to GFP present in all. The antisera from competitors A and B show more background reactivity in the E. coli control (lane A). Slight Contaminants Not run Competitor E Multiple peaks Contaminants n/a Competitor F Multiple peaks Contaminants n/a Competitor G Multiple peaks Clean n/a Some Background High Background Visit now to ind the products you need for your immunological research, or get in touch on info@nordicmubio.com n/a n/a

5 Human Immunoglobulins - offers the highest quality range of products for the study and analysis of human immunoglobulins. Isotyping in research In vaccine research, the isotype of the antibodies produced postvaccination can give information on the type of immune response induced. For example, stronger induction of an IgA response might be desirable for a vaccine targeting viral pathogens, inducing mucosal immunity. While higher levels of IgG2 are correlated with a Th1 (proinflammatory) response, and higher IgG1 is more characteristic of a Th2 (anti-inflammatory) response. Screening for antibodies developed against biologic drugs is a requirement of most clinical trials and post-approval monitoring. When a patient does develop an immune response it can be useful to determine both the specificity and antibody isotype of the response. Our highly validated product range spans mouse monoclonal antibodies against all isotypes of human immunoglobulins, including allotypes, and are available in multiple conjugates suitable for applications such as ELISA, flow cytometry, western blot and immunohistochemistry. We also have a range of highly purified human immunoglobulins, suitable for use as references and antigens. Determining the class (isotype) and subclass of an antibody is important for a variety of reasons, from selection of purification protocols to day-to-day immunological methods and clinical diagnostics. Different antibody classes and subclasses serve a variety of different functions within the body and can be indicative of disease states and immunological processes. For example, IgE is characteristic of allergic reactions. Each antibody isotype has unique effector functions, capable of binding different cellular receptors. Visit nordicmubio.com/products now to find the answer to all your human immunoglobulin needs, or get in touch on info@nordicmubio.com

6 Human Immunoglobulin Products Purified Human Immunoglobulin Cat. Number IgG HuIgG IgG subclasses standard Serum NOR-01 IgG1 lambda G1m(f) P 106 IgG4 kappa Gm4a P 182 IgG1 lambda G1m (a) P 210 IgG3 lambda G3m (g) P 289 IgG4 kappa Gm4b P 304 IgG2 lambda G2m n-) P 315 IgG3 kappa G3m(b) P 411 IgG2 lambda G2m (n+) P 532 IgA subclasses standard Serum NOR-04 Secretory IgA P 020 IgA1 lambda (dimer) P 444 IgA1 kappa, polymer P 430 IgA2 lambda A2m (1-, 2+) P 505 IgA2 (1+,2-) lambda P 237 Human IgE kappa P 050 Human Bence Jones kappa P 016 protein (Free light chain, kappa type) Human Bence Jones lambda P017 protein (Free light chain, lambda type) Available Mouse anti-human Cat. Number Conjugates* IgG1 MAHu/IgG1 B,H,F,T IgG2 (Fab subclass specific) MAHu/IgG2 B,H,F,T IgG2 (Fc subclass specific) MAHu/IgG2Fc B,H,F,T IgG3 (F(ab'')2 subclass specific) MAHu/IgG3 B,H,F,T IgG3 (G3m(U) allotype specific) MAHu/IgG3m(u) B,H,F,T IgG4 (pfc subclass specific) MAHu/IgG4 B,H,F,T IgG (class specific) MAHu/IgGc B,H,F,T IgM (class specific) MAHu/IgM B,H,F,T IgA1 MAHu/IgA1 B,H,F,T IgA2 MAHu/IgA2 B,H,F,T IgA2 (m2 allotype) MAHu/IgA2(m)2 B,H,F,T IgA1 + IgA2 MAHu/IgAc B,H,F,T - has developed an ever-expanding range of high quality immunological research tools, manufactured to strict ISO 9001 guidelines, to provide both reassurance and the best possible tools for your research. Visit nordicmubio.com/products now to find the answer to all your human immunoglobulin needs, or contact us: info@nordicmubio.com IgE MAHu/IgE B,H,F,T IgD MAHu/IgD B,H,F,T *B= Biotin, H=Horseradish peroxidase, F = FITC, T=TRITC Contact your local distributor here:

7 Veterinary Immunoglobulins - has developed an ever-expanding range of high quality immunological research tools, manufactured to strict ISO 9001 guidelines, to provide both reassurance and the best possible tools for your research. - is dedicated to helping veterinary scientists advance their understanding of animal immunology, helping to safeguard everything from domestic pets to livestock in health and disease. Our highly validated veterinary immunoglobulin products include a wide range of both host and target species, giving researchers the scope and choice they need. As in humans, animal immunoglobulins play a huge role in health and disease, with different antibody classes and subclasses indicative of different disease states or normal activity of immunological processes. This makes animal immunoglobulins of interest and importance to a huge range of research areas from infectious diseases to cancer research. We know your research needs reliable products that just work. And that s why we ve developed a portfolio of veterinary immunoglobulin products with a wealth of validation data, suitable for use as either primary or secondary antibodies and in a range of formats to suit your research needs. Whether your application is immunoprecipitation, ELISA or immunohistochemistry, if we list it as an application you can be sure that our products will work. Saving you not only money, but more importantly, time and precious sample. Together with our strict ISO 9001 manufacturing and quality control you can be sure each of our products will work time after time, lot after lot. Visit now to find the products you need for your immunological research, or get in touch on info@nordicmubio.com

8 Our 15 Top Selling Veterinary Immunoglobulin Products Our complete range of veterinary products includes reagents for research into: Cow, Dog, Chicken, Duck, Electric Eel, Cat, Goat, Guinea Pig, Hamster, Horse, Mouse, Rabbit, Rat, Sheep, Snake, Swine and many more. Anti IgG* Rabbit Goat RAG/IgG(H+L) B, F, H, T EL, ICC, IHC(P), DB, IB, IP Swine Mouse SwAM/IgG(H+L) F EL, ICC, IHC(F), IF Rabbit Swine RASw/IgG(H+L) B, F, H, T EL, ICC, IHC(P), DB, IB, IP, IF Rabbit Dog RAD/IgG(H+L) B, F, H, T EL, ICC, IHC(P), DB, IB, IP Goat Cow GAB/IgG(H+L) B, F, H EL, ICC, IHC(P), IHC(F), DB, IB, IP Goat Rabbit GAR/IgG(H+L) B, F, H, T EL, ICC, IHC(P), IHC(F), DB, IB, IP Rabbit Horse RAHo/IgG(H+L) B, F, H EL, ICC, IHC(P), IHC(F), DB, IB, IP Rabbit Cat RACa/IgG(H+L) B, F, H EL, ICC, IHC(F), DB, IB, IP, IF Rabbit Duck RADu/IgG(H+L) B, F, H EL, ICC, IHC(P), IHC(F), DB, IB, IP Rabbit Chicken RACh/IgG(H+L) B, F, H EL, ICC, IHC(P), DB, IB, IP, IF Anti IgD (Fc specific) Goat Mouse GAM/IgD(Fc) B, F, H, T EL, ICC, IHC(F), IP, IF Anti IgG1, IgG2a, IgG2b, IgG3, IgA, IgM* Rabbit Mouse RAM/Ig B, F, H, T EL, ICC, IHC(P), IHC(F), DB, IB, IP Applications Key: EL: ELISA ICC: Immunocytochemistry IHC(F): Immunohistochemistry (frozen) IHC(P): Immunohistochemistry (parrafin) DB: Dot blotting IB: Immunoblotting IP: Immunoprecipitation IF: (in)direct immunofluorescence Conjugate Key: B: Biotin H: Horseradish peroxidase F: FITC T: TRITC Anti IgG1, IgG2a, IgG2b, IgG3* Goat Mouse GAM/IgG(H+L) B, F, H, T EL, ICC, IHC(P), DB, IB, IP Anti serum proteins Rabbit Horse RAHo/TSP _ IP Anti Beta lactoglobulin Rabbit Cow RAB/BLgl _ IP Contact your local distributor here: *Heavy and Light chains