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Product Name
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Human Dual Interferon Gamma + Granzyme B Elispot Kit (Reagents for 10 x 96 Well Plates)
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Product type
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Kits
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Description
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Human Dual Interferon Gamma + Granzyme B Elispot Kit (Reagents for 10 x 96 Well Plates)
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Abreviews
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Research areas
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Kits/ Lysates/ Other >> Kits >> ELISA Kits >> ELISPOT Kits
Cancer >> Tumor immunology >> Cytokines >> Interferons
Stem Cells >> Signaling Pathways >> TGF beta >> Interferon gamma
Immunology >> Innate Immunity >> Cytokines >> Interferons
Cell Biology >> Apoptosis >> Extracellular Signals >> Granzymes
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Product overview
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The ELISPOT assay is designed to enumerate cytokine producing cells in a single cell suspension. This method has the advantage of requiring a minimum of in-vitro manipulations allowing cytokine production analysis as close as possible to in-vivo conditions in a highly specific way. This technique is designed to determine the frequency of cytokine producing cells under a given stimulation, and the follow-up of such frequency during a treatment and/or a pathological state. Elispot assay constitutes an ideal tool in the TH1 / TH2 response, vaccine development, viral infection monitoring and treatment, oncology, infectious diseases, autoimmune diseases and transplantation.
This Elispot assay is based on sandwich immuno-enzyme technology. Cell secreted cytokines or soluble molecules are captured by coated antibodies avoiding diffusion in supernatant, protease degradation or binding on soluble membrane receptors. After cell removal, the captured cytokines are revealed by tracer antibodies and appropriate conjugates.
The dual colour Elispot allows you to monitor the production of two cytokines simultaneously in the same well.
Principle of the Method
After cell stimulation, locally produced cytokines are captured by IFN gamma and Granzyme B specific monoclonal antibodies. After cell lysis, trapped cytokine molecules are revealed by a secondary anti-IFN gamma FITC conjugated antibody and a biotinylated anti-Granzyme B antibody. Those are in turn recognised by anti-FITC HRP and streptavidin-AP conjugates. PVDF-bottomed-well plates are then incubated first with AEC substrate buffer, washed and subsequently incubated with BCIP/NBT. Coloured red/brownish spots indicate IFN gamma production while Granzyme B is revealed by blue/purple spots.
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Components
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Kit Components |
Unit Size/Volume |
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| Capture antibody for Interferon gamma | 2 vials of 0.5 ml | | Capture antibody for Granzyme B | 2 vials of 0.5 ml | | FITC conjugated detection antibody for Interferon gamma | 2 vials lyophilised, resuspend each in 0.55mL | | Biotinylated detection antibody for Granzyme B | 2 vials lyophilised, resuspend each in 0.55mL | | Anti-FITC antibody HRP conjugate | 2 x 100 ul | | Streptavidin Alkaline Phosphatase conjugate | 2 x 50ul | | Bovine Serum albumin | 2 x 1 g | | Dry skimmed milk | 2 x 1 g | | 50 x concentrate AEC substrate buffer | 2 x 1 mL | | 10 x concentrate buffer for the preparation of AEC buffer | 2 x 5 mL | | Ready-to-use BCIP/NBT substrate buffer | 2 x 50 mL |
Note: For a detailed set of instructions please see the protocol.
Reagents and materials not provided: - 96 wells polysterene plate (eg. Nunc Maxisorp Cat#468667)
- Cell culture media
- CO2 incubator
- 70% ethanol
- Tween 20
- Phosphate buffered saline
- ELISPOT reading system.
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Storage instructions
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Store at 4°C. The kit is shipped at room temperature
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Species Reactivity
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Reacts with Human.
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Tested applications
(see key)
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ELISPOT
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Material safety datasheet (MSDS) for this product:
Dual Elispot Kit MSDS
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Relevance
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Interferon Gamma is produced by lymphocytes activated by specific antigens or mitogens. IFN-gamma, in addition to having antiviral activity, has important immunoregulatory functions. It is a potent activator of macrophages, it has antiproliferative effects on transformed cells and it can potentiate the antiviral and antitumor effects of the type I interferons.
Granzyme B is necessary for target cell lysis in cell-mediated immune responses. It cleaves after Asp. Seems to be linked to an activation cascade of caspases (aspartate-specific cysteine proteases) responsible for apoptosis execution. Cleaves caspase-3, -7, -9 and 10 to give rise to active enzymes mediating apoptosis.
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Alternative names Human Dual Interferon Gamma + Granzyme B Elispot Kit (Reagents for 10 x 96 Well Plates) (ab48465) |
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Database links
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The links below go to external sites and will open in a new browser window |
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At Abcam, we have one centralized database to hold all of our product information, so that everything we know about this Human Dual Interferon Gamma + Granzyme B Elispot Kit (Reagents for 10 x 96 Well Plates) is on this datasheet. But please do contact us if you would like any reassurance! |
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Search PubMed (MEDLINE) for references to Interferon Gamma + Granzyme B
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