Invitrogen™ Mouse IL-27 Uncoated ELISA Kit

Description

The Mouse Interleukin 27 (IL-27) 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 mouse IL-27. 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.

IL-27, a member of the IL-12 family, is a heterodimeric protein consisting of the p40-related protein Epstein-Barr virus-induced gene 3 (EBI3) non-covalently linked to an IL-12p35-related protein, p28 (also known as IL-30). IL-27 is produced by activated APCs and mature dendritic cells. IL-27 exerts its activities on NK cells and naive CD4+ T cells; mRNA expression analysis of IL-27 receptor components (WSX-1/TCCR and gp130) suggests that IL-27 may also target other cells, including mast cells and monocytes. Binding of IL-27 to WSX-1/gp130 activates JAK1, STAT1, and STAT3 and STAT1/3 phosphorylation. WSX-1/TCCR-deficient mice develop impaired Th1 responses and are more susceptible to infection with L. monocytogenes suggesting that Th1 responses require IL-27. Although activation of WSX-1 is required for the initiation of Th1 responses, it is not necessary for maintaining Th1 responses. IL-27 alone is not able to induce the differentiation of CD4+ T cells into IFN-γ-producing cells, suggesting a role for IL-27 as an initial activator of Th1 responses. An important effect of IL-27 in initiating Th1 responses is the induction of the Th1-specific transcription factor T-bet as well as the suppression of the Th2-specific transcription factor GATA-3. T-bet plays a critical role in Th1 differentiation by its ability to maintain IL-12Rβ2 expression following CD4+ T cell activation. Recent studies indicate that IL-27 has a potent antitumor activity. In vitro, IL-27 has been found to act directly on naive CD8 cells, generating CTL with enhanced granzyme B expression. In vivo, IL-27 has been reported to augment CTL activity, inhibit tumor growth, and induce complete regression of primary and metastatic neuroblastoma tumors.