Product nameHuman Interferon gamma + IL5 ELISPOT Kit (without plates)
See all Interferon gamma + IL5 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, cancerology, infectious diseases, autoimmune diseases and transplantation.
The 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 IL-5 specific monoclonal antibodies. After cell lysis, trapped cytokine molecules are revealed by a secondary anti-IFN gamma FITC conjugated antibody and a biotinylated anti-IL-5 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 IL-5 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-5 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-5 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
RelevanceMammalian Interferon gamma is mainly produced by T lymphocytes and NK cells. It is a pleiotropic cytokine involved in the regulation of nearly all phases of immune and inflammatory responses,including the activation, growth and differentiation of T cell, B cells, macrophages, NK cells and other cell types such as endothelial cells and fibroblasts. It has weak antiviral and antiproliferative activity, and poteniates the antiviral and anti tumor effects of IFN alpha / beta (type I interferon). It is upregulated by IL2, FGF basic, EGF and downregulated by vitamin D3 or DMN. Labile at pH 2. Interleukin 5 (IL5) is a cytokine produced primarily by activated T lymphocytes. It exists as an antiparallel disulfide linked homodimeric glycoprotein with 115 amino acid residues in each chain. Known also as EDF (eosinophil differentiating factor), functions predominantly as an eosinophilopoietic factor. Analysis of its crystal structure reveals a novel two domain structure, with each domain showing significant homology to the cytokine fold in GMCSF, MCSF, IL2, IL4 and growth hormone. Human and mouse IL5 have 70% amino acid sequence homology.
- B cell differentiation Factor 1
- Eosinophil differentiation factor
Our Abpromise guarantee covers the use of ab46655 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.|
ab46655 has not yet been referenced specifically in any publications.