Murine IL-4 ELISA Kit
The mIL-4 ELISA is an enzyme-linked immunosorbent assay for quantitative detection of murine Interleukin-4 in cell culture supernatants, murine serum, plasma or other body fluids.
IL-4 mediates ist function by binding to receptors expressed on target cells. The IL-4 receptors exhibit an affinity of approximately 10-10 M.
Receptors exist on freshly prepared B and T lymphocytes and macrophages, as well as on various cell lines including lymphoid cells, mast cell lines, a variety of other hematopoietic cell lines, fibroblasts and stromal cell lines. On T and B cells, receptors are present in low numbers (appr. 400), which are reported to be upregulated by IL-2 and IL-4.
An anti-mIL-4 monoclonal coating antibody is adsorbed onto microwells.
mIL-4 present in the sample or standard binds to antibodies adsorbed to the microwells; a biotin-conjugated monoclonal anti-mIL-4 antibody is added and binds to mIL-4 captured by the first antibody.
Following incubation unbound biotin conjugated antimIL-4 is removed during a wash step. Streptavidin-HRP is added and binds to the biotin conjugated anti-mIL-4. Following incubation unbound Streptavidin-HRP is removed during a wash step, and substrate solution reactive with HRP is added to the wells.
A coloured product is formed in proportion to the amount of mIL-4 present in the sample. The reaction is terminated by addition of acid and absorbance is measured at 450nm. A standard curve is prepared from seven mIL-4 standard dilutions and mIL-4 sample concentration determined.