ANGPTL6, Human, BioAssay(TM) ELISA Kit (Angiopoietin-related Protein 6, Angiopoietin-like Protein 6, Angiopoietin-related Growth Factor, Angiopoietin-related Protein 5, UNQ152/PRO178, AGF, ARP5)

Seven angiopoietin-like proteins (ANGPTLs) share the characteristic protein structure of the angiopoietin family (ANG), but differ in their inability to bind antiopoietin receptor, Tie-2. ANGPTL6 was originally named angiopoietin-related growth factor (AGF) having an N-terminal coiled-coil-like domain and a C-terminal fibrinogen-like domain, both of which are conserved in ANG. It is a circulating protein secreted by liver that induces angiogenesis by direct effect of epidermal ANGPTL6 on endothelial cells and proliferation of skin cells, and thereby promotes wound healing. 80% of Angptl6 -/- mice died at about embryonic day 13. The surviving null mice developed marked obesity, lipid accumulation in skeletal muscle and liver, and insulin resistance accompanied by reduced energy expenditure relative to controls. Conversely, mice with constitutive overexpression of ANGPTL6 showed leanness and increased insulin sensitivity resulting from increased energy expenditure, and were also protected from high-fat diet-induced obesity, insulin resistance, and non-adipose tissue steatosis. This assay is a sandwich ELISA for quantitative determination of human ANGPTL6 in biological fluids. A monoclonal antibody specific for ANGPTL6 has been precoated onto the 96-well microtiter plate. Standards and samples are pipetted into the wells for binding to the coated antibody. After extensive washing to remove unbound compounds, ANGPTL6 is recognized by the addition of a purified polyclonal antibody specific for ANGPTL6 (Detection Antibody). After removal of excess polyclonal antibody, HRP conjugated anti-rabbit IgG (Detector) is added. Following a final washing, peroxidase activity is quantified using the substrate 3,3',5,5'-tetramethylbenzidine (TMB). The intensity of the color reaction is measured at 450nm after acidification and is directly proportional to the concentration of ANGPTL6 in the samples. The assay range is 1.56-100ng/ml ANGPTL6/ml. The lowest level of ANGPTL6 that can be detected by this assay is 1.2ng/ml.\n\nSample Type:\nCell and tissue culture supernatants, urine, plasma, serum, as well as many other biological fluids.\n\nIntended Use:\nFor the quantitative determination of human ANGPTL6 in biological fluids.\n\nSensitivity:\n1.2ng/ml\n\nRange:\n1.56-100ng/ml ANGPTL6/ml\n\nSpecificity:\nMammalian\n\nTest Principle:\nThis assay is a sandwich ELISA for quantitative determination of human ANGPTL6 in biological fluids. A monoclonal antibody specific for ANGPTL6 has been precoated onto the 96-well microtiter plate. Standards and samples are pipetted into the wells for binding to the coated antibody. After extensive washing to remove unbound compounds, ANGPTL6 is recognized by the addition of a purified polyclonal antibody specific for ANGPTL6 (Detection Antibody). After removal of excess polyclonal antibody, HRP conjugated anti-rabbit IgG (Detector) is added. Following a final washing, peroxidase activity is quantified using the substrate 3,3',5,5'-tetramethylbenzidine (TMB). The intensity of the color reaction is measured at 450nm after acidification and is directly proportional to the concentration of ANGPTL6 in the samples.\n\nKit Components:\nPre-coated Microtiter Plate: 12x8 well strips\nWash Buffer (10X): 1x50ml\nDiluent (5X):1x50ml\nDetection Antibody: 1x12ml\nDetector 100X (HRP Conjugated anti-rabbit IgG): 1 vial\nHuman ANGPTL6 Standard (lyophilized): 200ng\nHuman ANGPTL6 QC Sample (lyophilized): 1 vial\nSubstrate Solution I (TMB): 1x6ml\nSubstrate Solution II (Peroxidase): 1x6ml\nStop Solution: 1x12ml\nPlate Sealers (plastic film): 3 each\n\nStorage and Stability:\nStore powder at 4 degrees C liquid at -20 degrees C. Store other components at 4 degrees C. Stable for at least 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.

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