General 8-Hydroxydeoxyguanosine ELISA Kit

A competitive enzyme-linked immunosorbent assay (cELISA) from Antibodies.com designed for the in vitro quantitative determination of 8-Hydroxydeoxyguanosine in serum, plasma or other biological fluids. The assay employs the competitive enzyme immunoassay technique on a 96-well microtiter plate pre-coated with 8-Hydroxydeoxyguanosine antigen. During incubation, analyte present in samples or standards competes with the immobilized antigen for binding sites on a Biotinylated Anti-8-Hydroxydeoxyguanosine Antibody; the more analyte present, the less antibody binds. After washing, an HRP-Avidin conjugate is added, and a TMB substrate is used to visualise the reaction, producing a blue product that changes to yellow upon addition of acidic stop solution. The intensity of the yellow colour is inversely proportional to the analyte concentration and is read at 450 nm on a colorimetric microplate reader.

The working range is 62.5–4000 pg/ml with a sensitivity of 18.5 pg/ml (described as <18.5 pg/ml). Total assay time is 2h 30m. Spike-and-recovery in serum averaged 91% (range 84–99%, n=5), in EDTA plasma 90% (range 82–98%, n=5) and in heparin plasma 93% (range 84–102%, n=5). Linearity across serial dilutions was as follows: serum (n=5) 72–106% at 1:2, 90–96% at 1:4, 84–104% at 1:8 and 82–101% at 1:16; EDTA plasma (n=5) 95–101%, 84–110%, 83–96% and 90–104%; heparin plasma (n=5) 74–103%, 90–93%, 93–105% and 81–106%. The kit includes a pre-coated 96-well microplate (12 x 8 well strips), lyophilized standard, detection solutions A (70 µl) and B (120 µl), 30X wash buffer (20 ml), sample dilution buffer (45 ml), TMB substrate (9 ml), stop solution (6 ml) and plate sealers. Components are stored at +4 °C or –20 °C as instructed and should not be used past the expiration date.