IL10 + IL2 Human Fluorospot Kit (Reagents for 10 x 96 tests, without plates) (ab48461)
- Product nameIL10 + IL2 Human Fluorospot Kit (Reagents for 10 x 96 tests, without plates)See all IL10 + IL2 kits ...
- Tests10 x 96 well plate
- Sample typeCell culture supernatant, Cell culture extracts
- Assay typeSandwich (qualitative)
- Assay durationMultiple steps standard assay
- 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, 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.
Principle of Method
After cell stimulation, locally produced cytokines are captured by IL-10 and IL-2 specific monoclonal antibodies. After cell lysis, trapped cytokine molecules are revealed by a secondary anti- IL-10 FITC conjugated antibody and a biotinylated anti-IL-2 antibody. Those are in turn recognised by anti-FITC green fluorescent dye and streptavidin-phycoerythrin conjugates. PVDF-bottomed-well plates are then read under a UV light beam. Green fluorescent spots indicate IL-10 production while IL-2 is revealed by red spots. Yellow spots will indicate dual cytokine producing cells.
- Tested applicationsFluorospot more details
- Storage instructionsStore at +4°C. Please refer to protocols.
Components 10 x 96 tests Anti-FITC antibody green fluorescence conjugate. 2 vials Biotinylated detection antibody for IL-2 (resuspend in 0.55mL). 2 x 0.55ml Bovine Serum albumin. 2 x 1g Capture antibody for IL-10 2 x 0.5ml Capture antibody for IL-2 2 x 0.5ml FITC conjugated detection antibody for IL-10(resuspend in 0.55mL). 2 x 0.55ml Fluorescence buffer 2 x 2.5ml Streptavidin-phycoerythrin conjugate. 2 vials
- Research Areas
- RelevanceIL2 is produced by T-cells in response to antigenic or mitogenic stimulation, this protein is required for T-cell proliferation and other activities crucial to regulation of the immune response. Can stimulate B-cells, monocytes, lymphokine-activated killer cells, natural killer cells, and glioma cells. IL10 inhibits the synthesis of a number of cytokines, including IFN-gamma, IL-2, IL-3, TNF and GM-CSF produced by activated macrophages and by helper T-cells.
- Cellular localizationSecreted
- Cytokine synthesis inhibitory factor
- Interleukin 10
- Interleukin 2
- T cell growth factor
Our Abpromise guarantee covers the use of ab48461 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
References for IL10 + IL2 Human Fluorospot Kit (Reagents for 10 x 96 tests, without plates) (ab48461)
ab48461 has not yet been referenced specifically in any publications.