Sirtuin 1 (human intracellular) ELISA Kit

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 Enzyme Linked-Immunosorbent Assay (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 450 nm 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 – 2 ng Sirtuin 1/ml and a detection limit of 30 pg/ml (based on adding two standard deviations to the mean value of the (50) zero standards).