Description
Human Immunoglobulin E (IgE) ELISpot Antibody Pair Kit is intended for those who want the flexibility of setting up their own ELISpot assay. This kit is designed for the quantitative detection and enumeration of cells secreting human IgE in a suspension of peripheral blood mononuclear cells (PBMCs). ELISpot is commonly used for evaluating immune responses, such as determining the frequency and isotype of circulating antibody-secreting cells (ASCs). Soluble IgE is secreted by IgE+ ASCs and contributes to host defense against parasitic infections as well as allergic responses. ELISpot is highly sensitive and can detect secreted analytes at a single-cell level. This assay is used in research of allergies, infectious diseases, and vaccines.
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 immunoglobulin. Secreted immunoglobulin is then captured throughout the culture period before the cells are washed away. Captured immunoglobulin is detected by the addition of a biotinylated detection antibody, followed by 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 total IgE-secreting cells in the sample and the amount of immunoglobulin secreted.
NOTE: This kit includes sufficient reagents for 4 x 96-well ELISpot plates. ELISpot plates (e.g. Millipore Sigma Catalog #MSIPS4W10/MAIPSWU10) and BCIP/NBT substrate (e.g. Catalog #200-1016) are required for use with this kit and are available for purchase separately.
This kit is supplied in two boxes (Catalog #200-1008/200-1009).
