Interleukin 6 (IL-6),also known as interferon-beta 2,26kD protein and B cell stimulatory factor-2 (BSF-2),is a pleiotropic alpha-helical cytokine that plays important roles in acute phase reactions,inflammation,hematopoiesis,bone metabolism,and cancer progression. IL-6 activity is essential for the transition from acute inflammation to either acquired immunity or chronic inflammatory disease. It is secreted by multiple cell types as a 22kD - 28kD phosphorylated and variably glycosylated molecule. Mature human IL-6 is 183aa in length and shares 41% aa sequence identity with mouse and rat IL-6. Alternate splicing generates several isoforms with internal deletions,some of which exhibit antagonistic properties.Sample Type:Cell culture supernatants,serum,plasma,and tissueIntended Use:The Human IL-6 BioAssay ELISA Kit is an enzyme-linked immunosorbent assay for the quantitative detection of Human IL-6 concentrations in cell culture supernatants,serum,plasma,and tissue.Sensitivity:1pg/mlRange:3.9-250pg/mlSpecificity:Recognizes human IL-6. There was no detectable cross reactivity with any of the tested proteins following IL-1,2,3,4,5,8,10,12,TNF,VEGF,TGF,IFN,and sICAM.Sample Volume:100ul/wellTest Principle:This assay employs the Sandwich immunoassay technique. An anti-h IL-6 monoclonal coating antibody is adsorbed onto microwells. IL-6 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-6 antibody is added and binds to IL-6 captured by the first antibody. Following incubation unbound Biotinylated anti-h IL-6 antibody is removed during a wash step. A Streptavidin-HRP is added and binds to Biotinylated anti-h IL-6 antibody. Following incubation unbound Streptavidin-HRP is removed during a wash step. A colored product is formed in proportion to the amount of IL-6 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-6 standard dilutions and IL-6 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.