Murine IL-5 ELISA Kit
The mIL-5 ELISA is an enzyme-linked immunosorbent assay for quantitative detection of murine Interleukin-5 in cell culture supernatants, murine serum, plasma or other body fluids.
Mouse IL-5 has been described as T-cell replacing factor and as B-cell growth factor II based on its induction of proliferation and Ig secretion by activated B cells. Several other activities have furthermore been attributed to this cytokine, including induction of growth and differentiation of eosinophils from bone marrow stew cells, induction of mIL-2R expression on both normal splenic and activated B cells and enhancements of IgA secretion by LPS-stimulated B-cells. m-IL-5 has been shown to be produced exclusively by TH2 subset of mouse TH clones. The spectrum of biological activities of mIL-5, on B cells as well as other cell types suggests that mIL-5 plays an important functional role in the activities of TH2 cells. mIL-5 is a heavily glycosylated glycorprotein with native m.w. of 45.000 to 60.000, with multiple subunits of between m.w. 24.000 and 28.000 under reducing conditions.
An anti-mIL-5 monoclonal coating antibody is adsorbed onto microwells.
mIL-5 present in the sample or standard binds to antibodies adsorbed to the microwells; a biotin-conjugated monoclonal anti-mIL-5 antibody is added and binds to mIL-5 captured by the first antibody.
Following incubation unbound biotin conjugated anti-mIL5 is removed during a wash step. Streptavidin-HRP is added and binds to the biotin conjugated anti-mIL-5. 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-5 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-5 standard dilutions and mIL-5 sample concentration determined.