Adiponectin (also known as Adipocyte complement-related protein of 30kD,Acrp30) is an adipocyte-derived protein with wide ranging paracrine and endocrine effects on metabolism and inflammation. It promotes adipocyte differentiation,fatty acid catabolism,and insulin sensitivity and is negatively correlated with obesity,type 2 diabetes,and atherogenesis. Adiponectin associates into 70kD noncovalently-linked trimers that are disulfide-linked into medium molecular weight hexamers and then into 300kD high molecular weight oligomers.Sample Type:Cell culture supernatants,serum,plasma,and tissueIntended Use:The Human Adiponectin BioAssay ELISA Kit is an enzyme-linked immunosorbent assay for the quantitative detection of Human Adiponectin concentrations in cell culture supernatants,serum,plasma,and tissue.Sensitivity:50pg/mlRange:156-10000ng/mlSpecificity:Recognizes human Adiponectin.Sample Volume:100ul/wellTest Principle:This assay employs the Sandwich immunoassay technique. An anti-h Adiponectin monoclonal coating antibody is adsorbed onto microwells. Adiponectin 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 Adiponectin antibody is added and binds to Adiponectin captured by the first antibody. Following incubation unbound Biotinylated anti-h Adiponectin antibody is removed during a wash step. A Streptavidin-HRP is added and binds to Biotinylated anti-h Adiponectin antibody. Following incubation unbound Streptavidin-HRP is removed during a wash step. A colored product is formed in proportion to the amount of Adiponectin 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 Adiponectin standard dilutions and Adiponectin 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.