Interleukin 10 (IL-10),initially designated cytokine synthesis inhibitory factor (CSIF),was originally identified as a product of mouse T helper (Th) 2 clones that inhibited cytokine production by Th1 clones. The human ortholog of mouse IL-10 was subsequently cloned by cross-hybridization. Human and mouse IL-10 are 81% identical at the nucleotide and amino acid level. IL-10 is the prototypic member of the IL-10 cytokine family,including IL-10,IL-19,IL-20,IL-22 (IL-TIF),IL-24 and IL-26. A number of viruses,including Epstein-Barr virus (EBV) and human cytomegalovirus (HCMV),also encode viral members of the IL-10 family. Human IL-10 has cross-species actvity and is active on mouse cells. Mouse IL-10 is species-specific and does not act on human cells.Sample Type:Cell culture supernatants,serum,plasma,and tissueIntended Use:The Human IL-10 BioAssay ELISA Kit is an enzyme-linked immunosorbent assay for the quantitative detection of Human IL-10 concentrations in cell culture supernatants,serum,plasma,and tissue.Sensitivity:7pg/mlRange:15.625-1000pg/mlSpecificity:Recognizes human IL-10.Sample Volume:100ul/wellTest Principle:This assay employs the Sandwich immunoassay technique. An anti-h IL-10 monoclonal coating antibody is adsorbed onto microwells. IL-10 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-h IL-10 antibody is added and binds to IL-10 captured by the first antibody. Following incubation unbound Biotinylated anti-h IL-10 antibody is removed during a wash step. A Streptavidin-HRP is added and binds to Biotinylated anti-h IL-10 antibody. Following incubation unbound Streptavidin-HRP is removed during a wash step. A colored product is formed in proportion to the amount of IL-10 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-10 standard dilutions and IL-10 sample concentration determined.Kit Components:Aluminium pouches with a Microwell Plate: 8x12Standard: 2xVialBiotinylated antibody: 2xVialStreptavidin-HRP: 2xVialAssay Buffer (25ml/Vial): 2xVialWash Buffer Concentrate 20x (30ml/Vial): 1xVialTMB Substrate Solution: 1x12mlStop Solution (12ml/Vial): 1xVialPlate Covers- Adhesive strips: 3xstripsInstruction: 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.