IL-17A ELISpot Assay Kit

$350.00$1,550.00

This IL-17A ELISpot Assay Kit is designed to determine the frequency of IL-17A producing cells under a given stimulation and the comparison of such frequency against a specific treatment or pathological state. The Eagle Biosciences IL-17A ELISpot Assay Kit is for research use only and not to be used for diagnostic procedures.

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IL-17A ELISpot Assay Kit

The IL-17A ELISpot Assay Kit is for research use only.

Incubation Time: 18hrs – 23hrs
Sample Size: 100 µl


Assay Principle

The ELISpot is a highly specific immunoassay for the analysis of cytokine and other soluble molecule production and secretion from T-cells at a single cell level in conditions closely comparable to the in-vivo environment with minimal cell manipulation. This technique is designed to determine the frequency of cytokine producing cells under a given stimulation and the comparison of such frequency against a specific treatment or pathological state. The ELISpot assay constitutes an ideal tool in the investigation of Th1 / Th2 responses, vaccine development, viral infection monitoring and treatment, cancerology, infectious disease, autoimmune diseases and transplantation. Utilizing sandwich immuno-enzyme technology, ELISpot assays can detect both secreted cytokines and single cells that simultaneously produce multiple cytokines. 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. A capture antibody highly specific for the analyte of interest is coated to the wells of a PVDF bottomed 96 well microtiter plate either during kit manufacture or in the laboratory. The plate is then blocked to minimize any non-antibody dependent unspecific binding and washed. Cell suspension and stimulant are added and the plate incubated allowing the specific antibodies to bind any analytes produced. Cells are then removed by washing prior to the addition of Biotinylated detection antibodies which bind to the previously captured analyte. Enzyme conjugated streptavidin is then added binding to the detection antibodies. Following incubation and washing substrate is then applied to the wells resulting in colored spots which can be quantified using appropriate analysis software or manually using a microscope.


Related Products

IL-17 ELISA Assay Kit
IL-17A/F ELISpot Assay Kit
IL-17F ELISpot Assay Kit

Additional Information

Assay Background


Interleukin-17 (IL-17, or IL-17A) is the founding member of a group of cytokines called the IL-17 family. IL-17A was originally identified as a transcript from a rodent T-cell hybridoma by Rouvier et al. in 1993. IL-17A is involved in inducing and mediating proinflammatory responses, commonly associated with allergic responses and induces the production of many other cytokines (such as IL-6, G-CSF, GM-CSF, IL-1β, TGF-β, TNFα), chemokines (including IL-8, GRO-α and MCP-1) and prostaglandins (e.g. PGE2) from many cell types (fibroblasts, endothelial cells, epithelial cells, keratinocytes and macrophages). IL-17A function is also essential to a subset of CD4+ T-Cells called T helper 17 (Th17) cells.

ELISpot Assay Kit Procedure

Documents

Product Manual