Product nameHuman Interferon Gamma + IL2 ELISPOT Kit (without plates)
See all Interferon Gamma + IL2 kits
Sample typeCell culture supernatant, Cell culture extracts
Assay typeSandwich (qualitative)
Assay durationMultiple steps standard assay
Species reactivityReacts with: Human
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.
After cell stimulation, locally produced cytokines are captured by IFN gamma and IL2 specific monoclonal antibodies. After cell lysis, trapped cytokine molecules are revealed by a secondary anti-IFN gamma FITC conjugated antibody and a biotinylated anti-IL2 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 IL2 is revealed by blue/purple spots.
Tested applicationsSuitable for: ELISpotmore details
Storage instructionsStore at +4°C. Please refer to protocols.
Components 5 x 96 tests 10 x 96 tests 15 x 96 tests 20 x 96 tests 10 x concentrate buffer for the preparation of AEC buffer 1 x 5ml 2 x 5ml 3 x 5ml 4 x 5ml 50 x concentrate AEC substrate buffer 1 x 1ml 2 x 1ml 3 x 1ml 4 x 1ml Anti-FITC antibody HRP conjugate 1 x 100µl 2 x 100µl 3 x 100µl 4 x 100µl Bovine Serum Albumin 1 x 1g 2 x 1g 3 x 1g 4 x 1g Human IFN Gamma Capture antibody 1 x 500µl 2 x 500µl 3 x 500µl 4 x 500µl Human IL-2 Capture antibody 1 x 500µl 2 x 500µl 3 x 500µl 4 x 500µl IFNγ FITC conjugated detection antibody 1 vial 2 vials 3 vials 4 vials IL-2 Biotinylated detection antibody 1 vial 2 vials 3 vials 4 vials Ready-to-use BCIP/NBT substrate buffer 1 x 50ml 2 x 50ml 3 x 50ml 4 x 50ml Streptavidin - Alkaline Phosphatase conjugated 1 x 50µl 2 x 50µl 3 x 50µl 4 x 50µl
RelevanceIFN-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.
- IFN gamma
Our Abpromise guarantee covers the use of ab48447 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ELISpot||Use at an assay dependent dilution.|
ab48447 has not yet been referenced specifically in any publications.