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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, cancerology, infectious diseases, autoimmune diseases and transplantation. 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 a specific monoclonal antibody. After cell lysis, trapped cytokine molecules are revealed by a secondary biotinylated detection antibody, which is in turn recognised by streptavidin conjugated to alkaline phosphatase. PVDF-bottomed-well plates are then incubated with BCIP/NBT substrate. Colored "purple" spots indicate cytokine production by individual cells.
|Components||1 x 96 tests||5 x 96 tests|
|Bovine Serum Albumin||1 x 200mg||1 x 1g|
|Human IL17 Biotinylated detection antibody||1 vial||1 vial|
|Human IL17 Pre-coated 96-well PDVF-bottomed plates||1 unit||5 units|
|Ready-to-use BCIP/NBT substrate buffer||1 x 10ml||1 x 50ml|
|Streptavidin - Alkaline Phosphatase conjugated||1 x 10µl||1 x 50µl|
Our Abpromise guarantee covers the use of ab83723 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 concentration.|
ab83723 has not yet been referenced specifically in any publications.