Interferon gamma + IL4 Human Elispot Kit (Reagents for 5 x 96 Well Plates) (ab46648)
- Product nameInterferon gamma + IL4 Human Elispot Kit (Reagents for 5 x 96 Well Plates)See all Interferon gamma + IL4 kits ...
- Tests5 x 96 well plate
- Sample typeCell culture supernatant, Cell culture extracts
- Assay typeELISPOT
- Species reactivityReacts with: Human
- Product overview
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, cancerology, 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 IL4 specific monoclonal antibodies. After cell lysis, trapped cytokine molecules are revealed by a secondary anti-IFN gamma FITC-conjugated antibody and a biotinylated anti-IL4 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 IL4 is revealed by blue/purple spots.
- Tested applicationsMeasuring secreted proteins from cells more details
- Storage instructionsStore at +4°C. Please refer to protocols.
Components 5 x 96 tests 10 x concentrate buffer for the preparation of AEC buffer 1 x 5ml 50 x concentrate AEC substrate buffer 1 x 1ml Anti-FITC antibody HRP conjugate 1 x 100µl Bovine Serum albumin 1 x 1g Human IFNγ Capture antibody 1 x 0.5ml IFNγ FITC conjugated detection antibody 1 x 0.55ml IL-4 Biotinylated detection antibody 1 x 0.55ml IL-4 Capture antibody 1 x 0.5ml Ready-to-use BCIP/NBT substrate buffer 1 x 50ml Streptavidin - Alkaline Phosphatase conjugated 1 x 50µl
- IFN gIFN gammaIl 4Immune interferon interferon gammainterleukin 4
Our Abpromise guarantee covers the use of ab46648 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
References for Interferon gamma + IL4 Human Elispot Kit (Reagents for 5 x 96 Well Plates) (ab46648)
ab46648 has not yet been referenced specifically in any publications.