SIRT1, Human, BioAssay(TM) ELISA Kit (NAD-dependent Protein Deacetylase Sirtuin-1, hSIRT1, Regulatory Protein SIR2 Homolog 1, SIR2-like Protein 1, hSIR2, SIR2L1)

Sirtuin 1 is the human ortholog for the yeast Sir2 (silent information regulator 2) protein regulating epigenetic gene silencing as a possible antiaging effect. Sirtuin 1 is an NAD(+)-dependent histone deacetylase, which deacetylate lysines 9 and 14 of histone H3 and lysine-16 of histone H4, involved in various cellular functions such as transcription, energy sensing, and differentiation. Sirtuin1 plays an important role in regulating adipogenesis via repression of PPAR and the gluconeogenic/glycolytic pathways in liver in response to fasting signals through the transcriptional coactivator PGC1A deacetylated at specific lysine residues in an NAD(+)-dependent manner. This assay is a sandwich ELISA for quantitative determination of human Sirtuin 1 in cells. A monoclonal antibody specific for Sirtuin 1 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, Sirtuin 1 is recognized by the addition of a purified polyclonal antibody specific for Sirtuin 1 (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 Sirtuin 1 in the samples. This ELISA is specific for the measurement of natural and recombinant human Sirtuin 1. It does not cross-react with human Sirtuin 2, human Sirtuin 5, human Sirtuin 6, human adiponectin, human resistin, human RBP4, human vaspin, human progranulin, human GPX3, human FTO, human Nampt, human leptin, mouse FTO, mouse Nampt. The assay range is 0.031-2ng Sirtuin 1/ml and a detection limit of 30pg/ml (based on adding two standard deviations to the mean value of the (50) zero standards).\n\nSample Type:\nCell lysates.\n\nIntended Use:\nFor the quantitative determination of human Sirtuin 1 in cells.\n\nSensitivity:\n30pg/ml \n\nRange:\n0.031-2ng Sirtuin 1/ml\n\nSpecificity:\nMammalian\n\nTest Principle:\nThis assay is a sandwich ELISA for quantitative determination of human Sirtuin 1 in cells. A monoclonal antibody specific for Sirtuin 1 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, Sirtuin 1 is recognized by the addition of a purified polyclonal antibody specific for Sirtuin 1 (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 Sirtuin 1 in the samples.\n\nKit Components:\nPre-coated Microtiter Plate: 12x8 well strips\nWash Buffer (10X): 1x50ml\nDiluent (5X):1x50ml\nLysis Buffer (10X): 1x12ml\nDetection Antibody: 1x12ml\nDetector 100X (HRP conjugated anti-IgG): 1x150ul\nHuman Sirtuin 1 Standard (lyophilized): 4ng\nHuman Sirtuin 1 QC Sample (lyophilized): 1 vial\nSubstrate Solution (TMB): 1x12ml\nStop Solution: 1x12ml\nPlate Sealers: 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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