Principle of Assay
The Mouse IL-4 ELISpot Kit (A334662) employs the double-antibody sandwich ELISpot technique for the detection of IL-4-secreting cells from mouse lymphocytes. Sample lymphocytes are isolated from mouse spleen tissue by mechanical grinding and plated into a 96-well microplate pre-coated with anti-IL-4 capture antibody. Upon stimulation with a cell stimulant, IL-4 secreted by individual cells is captured by the immobilized antibody. A biotinylated anti-IL-4 detection antibody, which also binds the captured cytokine, and Streptavidin-HRP, which binds the biotinylated detection antibody, are then added, and the plate is incubated. Following incubation, unbound detection antibody and Streptavidin-HRP are removed by washing, and AEC substrate solution is added. Colored spots develop at the sites of cytokine secretion, which can be quantified using an ELISpot reader. This ready-to-use kit includes pre-coated plates, detection antibody, Streptavidin-HRP, AEC coloring system, dilution buffer, cell stimulant, and washing buffer. For research use only. Not for diagnostic purposes.