Stemcell, 200-1003, Human IL-1 beta ELISpot Kit

Human interleukin-1 beta (IL-1β) ELISpot Kit is designed for the quantitative detection and enumeration of cells secreting human IL-1β in a suspension of peripheral blood mononuclear cells (PBMCs). ELISpot is commonly used for evaluating specific immune responses, such as determining the frequency of activated cells in response to stimulation. IL-1β is a pro-inflammatory cytokine secreted by monocytes and dendritic cells in response to infection. ELISpot is highly sensitive and can detect secreted analytes at a single-cell level. These assays are used in research of infectious diseases, cancers, allergies, and autoimmune diseases.

The assay is based on a modified version of the sandwich ELISA method, in which cells are added to ELISpot plates pre-coated with capture antibodies specific for the cytokine. The cytokine is then captured throughout the culture period and is detected by the addition of a biotinylated detection antibody, followed by streptavidin-alkaline phosphatase (SA-ALP), which binds to the biotinylated antibody. Addition of the chromogenic enzyme substrate 5-bromo-4-chloro-3-indolyl-1-phosphate (BCIP)/nitroblue tetrazolium (NBT), results in the formation of colored spots. The number, size, and intensity of these spots are directly proportional to the frequency of cytokine-secreting cells in the sample and the amount of analyte secreted.