Human Sirtuin 5 ELISA Kit

This sandwich ELISA kit from Antibodies.com is designed for the in vitro quantitative determination of human SIRT5 in tissue homogenates and other biological fluids. The assay is run on a pre-coated 96-well microplate arranged as 12 x 8 well strips, with a total assay time of 4 hours 30 minutes. Standards and test samples are added to the wells, where SIRT5 is captured by the immobilized antibody. A Biotinylated Anti-SIRT5 Antibody is then added, followed by an HRP-Avidin conjugate. The HRP enzymatic reaction is visualised using TMB substrate, producing a blue product that turns yellow upon addition of acidic stop solution. Absorbance is read at 450 nm in a colorimetric microplate reader and the SIRT5 concentration is calculated against the standard curve.

The kit has a stated measuring range of 0.156-10 ng/ml and a sensitivity of 0.067 ng/ml. Recovery has been validated in serum (n=5, average 91%, range 80-102%), EDTA plasma (n=5, average 90%, range 81-100%), and heparin plasma (n=5, average 84%, range 80-89%). Dilution linearity has been evaluated at 1:2, 1:4, 1:8, and 1:16 dilutions across the same sample types, with recoveries generally in the 74-108% range. Kit components include the pre-coated 96-well microplate, lyophilised standard (2 vials), Detection Solution A (120μl), Detection Solution B (120µl), 30X Wash Buffer (20ml), Sample Dilution Buffer (45ml), TMB Substrate (9ml), Stop Solution (6ml), and plate sealers (5 adhesive strips), with storage temperatures of +4°C or -20°C as specified per component.