The Human Interleukin 12 p70 (IL-12 p70) Uncoated ELISA Kit contains pre-matched antibody pairs, and reagents for performing quantitative enzyme linked immunosorbent assays (ELISA) to detect and quantify protein levels of human IL-12 p70. Wash Buffer and Stop Solution are needed to complete the ELISA reaction and are sold separately.
Principle of the method
ELISAs are designed to measure the amount of the target bound between a matched antibody pair. A target-specific antibody is coated to the bottom of the wells of a microplate, which is an overnight process. Samples, standards, or controls are then added into these wells and bind to the immobilized (capture) antibody. A sandwich is formed by the addition of the second (detector) antibody, a substrate solution is added that reacts with the enzyme-antibody-target complex to produce measurable signal. The intensity of this signal is directly proportional to the concentration of target present in the original specimen.
Interleukin-12 (IL-12) is a heterodimeric 70 kDa cytokine composed of two covalently linked, glycosylated chains with molecular weights of 40kD (p40) and 35-kD (p35). IL-12 is mainly produced by monocytes, macrophages, and dendritic cells in response to bacterial products such as lipopolysaccharides (LPS), to intracellular pathogens or upon interaction with activated T cells. IL-12 was originally discovered because of its ability to induce interferon-gamma (IFN-g) production, cell proliferation, and cytotoxicity mediated by natural killer cells and T cells. Studies have established that IL-12 also plays a key role in the development of Th1 responses, leading to IFN-g and IL-2 production. These cytokines can in turn promote T-cell responses and macrophage activation. Recombinant mouse IL-12 p70 is produced in baculovirus-infected insect cells as an authentic heterodimer of precursor p35 and p40 subunits using a dual promoter expression system. IL-12 p70 is distinct from other available forms of the protein in that it is expressed as a true heterodimer, as opposed to a single-chain, pseudo-heterodimer in which the subunits are joined by an artificial linker. The responses of lymphocytes to this cytokine are mediated by the activator of transcription protein STAT4. Nitric oxide synthase 2A (NOS2A/NOS2) is found to be required for the signaling process of this cytokine in innate immunity. Human and mouse IL-12 p40 share about 70% amino acid sequence homology.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.