Description
Principle of the Assay::The Rat IL-12 p40 ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of Rat IL-12 p40 in Cell Culture Supernatants, Serum, Plasma, Tissue Homogenates. This assay employs an antibody specific for Rat IL-12 p40 coated on a 96-well plate. Standards and samples are pipetted into the wells and IL-12 p40 present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-Rat IL-12 p40 antibody is added. After washing away unbound biotinylated antibody, HRP-conjugated streptavidin is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of IL-12 p40 bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.
Background/Introduction: Interleukin-12 (IL12; formerly NKSF, for natural killer cell stimulatory factor, or CLMF, for cytotoxic lymphocyte maturation factor) is a novel cytokine cloned from B-cell lines. IL-12 is a heterodimeric molecule composed of p35 and p40 subunits. The larger 40-kDa subunit (p40) is a member of the cytokine receptor family, and the smaller 35-kDa subunit (p35) is related to IL6 and GCSF. Both IL-12 p40 (-/-) and p35 (-/-) mice fail to produce IL-12 p70 heterodimer. Interleukin (IL)-12 has been cloned on the basis of its ability to activate natural killer (NK) cells and promote the development of cytolytic T cells. With further understanding of its activities, IL-12 has emerged as an important cytokine, affecting both immune and hematologic functions. It has been shown to be necessary for the T cell independent induction of interferon (IFN)-gamma, critical for the initial suppression of bacterial and parasitic infection; for the development of a Th1 response, critical for effective host defense against intracellular pathogens; and for the activation of differentiated T lymphocytes of both CD4+ and CD8+ phenotype. The subunits map to different chromosomes: p40 (IL12B) to 5q31-q33 and p35 (IL12A) to 3p12-3q13.2