Vaspin, Human, BioAssay(TM) ELISA Kit (Serpin A12, OL-64, Visceral Adipose Tissue-derived Serine Protease Inhibitor, Visceral Adipose-specific Serpin, SERPINA12)

Vaspin, designated as visceral adipose tissue-derived serpin, is a serpin whose expression is restricted to visceral adipose tissue. Vaspin seems to retain the serpin signature conformation fold consisting of three b-sheets and nine a-helices as well as a reactive loop site that interacts with its cognate serine protease unidentified hitherto. Vaspin is remarkably upregulated at high-fat high-sucrose (HFHS) diet and at multiple metabolic dysfunctions like obesity or insulin resistance. Administration of thiazolidinedione (TZD) caused a significant induction of serum vaspin in mice. These facts imply that upregulation of vaspin may be a compensatory response to antagonize the action of other unknown proteases upregulated in obesity or insulin resistance. Indeed, administration of recombinant vaspin to obese mice fed with HFHS chow improved glucose tolerance and insulin sensitivity. Vaspin seems to directly act on white adipose tissue. This assay is a sandwich ELISA for quantitative determination of human vaspin in biological fluids. A monoclonal antibody specific for vaspin 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, vaspin is recognized by the addition of a purified polyclonal antibody specific for vaspin (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 vaspin in the samples. The assay range is 0.016-1ng/ml Vaspin/ml. The lowest level of vaspin that can be detected by this assay is 12pg/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 vaspin in biological fluids.\n\nSensitivity:\n12pg/ml\n\nRange:\n0.016-1ng/ml Vaspin/ml\n\nSpecificity:\nMammalian\n\nTest Principle:\nThis assay is a sandwich ELISA for quantitative determination of human vaspin in biological fluids. A monoclonal antibody specific for vaspin 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, vaspin is recognized by the addition of a purified polyclonal antibody specific for vaspin (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 vaspin 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): 1x150ul\nHuman Vaspin Standard (lyophilized): 2ng\nHuman Vaspin QC Sample (lyophilized): 1 vial\nSubstrate Solution (TMB): 1x12ml\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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