Sirtuin 2 (human intracellular) ELISA Kit

Sirtuin 2 is a NAD-dependent histone deacetylase (39.5 kD) largely localized at the cytoplasm where it is physically associated with microtubules. SIRT2 was found to deacetylate lys40 of alpha-tubulin both in vitro and in vivo and colocalized and interacted in vivo with HDAC6, another tubulin deacetylase. It has been also shown that Sirtuin 2 is a regulator of mitotic progression that acts downstream from CDC14B in a pathway regulating mitotic exit or subsequent cytokinesis. This assay is a sandwich Enzyme Linked-Immunosorbent Assay (ELISA) for quantitative determination of human Sirtuin 2 in cells. A monoclonal antibody specific for Sirtuin 2 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 2 is recognized by the addition of a purified polyclonal antibody specific for Sirtuin 2 (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 2 in the samples. This ELISA is specific for the measurement of natural and recombinant human Sirtuin 2. It does not cross-react with human Sirtuin 1, 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 0.125 – 8 ng Sirtuin 2/ml and a detection limit of 80 pg/ml (based on adding two standard deviations to the mean value of the (50) zero standards).