Murine IL-12 ELISA Kit
The mIL-12 ELISA is an enzyme-linked immunosorbent assay for quantitative detection of murine Interleukin-12 in cell culture supernatants, murine serum, plasma or other body fluids.
Interleukin-12 (IL-12) is a pleiotropic cytokine, formerly termed cytotoxic lymphocyte maturation factor (CLMF) or natural killer cell stimulatory factor (NKSF) which is produced primarily by stimulated macrophages. IL-12 has been shown to be a proinflammatory cytokine produced by phagocytic cells, B cells, and other antigen – presenting cells that modulate adaptive immune responses by favoring the generation of T-helper type 1 cells.
An anti-mIL-12 monoclonal coating antibody is adsorbed onto microwells.
mIL-12 present in the sample or standard binds to antibodies adsorbed to the microwells; a biotin-conjugated monoclonal anti-mIL-12 antibody is added and binds to mIL-12 captured by the first antibody
Following incubation unbound biotin conjugated anti-mIL12 is removed during a wash step. Streptavidin-HRP is added and binds to the biotin conjugated anti-mIL-12. 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-12 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-12 standard dilutions and mIL-12 sample concentration determined.