IL-13 is a 17kD immunoregulatory cytokine that plays a key role in the pathogenesis of allergic asthma and atopy. It is secreted by Th1 and Th2 CD4+ T cells,NK cells,visceral smooth muscle cells,eosinophils,mast cells,and basophils. IL-13 circulates as a monomer with two internal disulfide bonds that contribute to a bundled four alpha-helix configuration. Mature human IL-13 shares 57%,59%,and 94%aa sequence identity with mouse,rat,and rhesus IL-13,respectively. Despite the low homology,it exhibits cross-species activity between human,mouse,and rat. IL-13 has diverse activities on numerous cell types. On macrophages,IL-13 suppresses the production of proinflammatory cytokines and other cytotoxic substances. On B cells,IL-13 induces immunoglobulin class switching to IgE,upregulates the expression of MHC class II,CD71,CD72,and CD23,and costimulates proliferation. IL-13 upregulates IL-6 while downregulating IL-1 and TNF-alpha production by fibroblasts and endothelial cells. IL-13 binds with low affinity to IL-13 R alpha 1,triggering IL-13 R alpha 1 association with IL-4 R alpha. This high affinity receptor complex also functions as the type 2 IL-4 receptor complex. Additionally,IL-13 binds with high affinity to IL-13 R alpha 2 which is expressed intracellularly,on the cell surface,and as a soluble molecule. IL-13 R alpha 2 regulates the bioavailability of both IL-13 and IL-4 and is overexpressed in glioma and several bronchial pathologies. Compared to wild type IL-13,the atopy-associated R110Q variant of IL-13 elicits increased responsiveness from eosinophils that express low levels of IL-13 R alpha 2.Sample Type:Cell culture supernatants,serum,plasma,and tissueIntended Use:The Mouse IL-13 BioAssay ELISA Kit is an enzyme-linked immunosorbent assay for the quantitative detection of Mouse IL-13 concentrations in cell culture supernatants,serum,plasma,and tissue.Sensitivity:4pg/mlRange:7.8-500pg/mlSpecificity:Recognizes mouse IL-13.Sample Volume:100ul/wellTest Principle:This assay employs the Sandwich immunoassay technique. An anti-m IL-13 monoclonal coating antibody is adsorbed onto microwells. IL-13 present in the sample or standard binds to antibodies adsorbed to the microwells. Following incubation unbound sample or standard are removed during a wash step. a Biotinylated anti-m IL-13 antibody is added and binds to IL-13 captured by the first antibody. Following incubation unbound Biotinylated anti-m IL-13 antibody is removed during a wash step. A Streptavidin-HRP is added and binds to Biotinylated anti-m IL-13 antibody. Following incubation unbound Streptavidin-HRP is removed during a wash step. A colored product is formed in proportion to the amount of IL-13 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 IL-13 standard dilutions and IL-13 sample concentration determined.Kit Components:Aluminium pouches with a Microwell Plate: 8x6Standard: 1xVialBiotinylated antibody: 1xVialStreptavidin-HRP: 1xVialAssay Buffer (25ml/Vial): 1xVialWash Buffer Concentrate 20x (30ml/Vial): 1xVialTMB Substrate Solution: 1x6mlStop Solution (12ml/Vial): 1xVialPlate Covers- Adhesive strips: 2xstripsInstruction: 1xStorage and Stability:Store powder at 4°C liquid at -20°C. Store other components at 4°C. Stable for at least 6 months For maximum recovery of product,centrifuge the original vial after thawing and prior to removing the cap.