DNA Damage ELISA Kit

The DetectX^® DNA Damage ELISA Kit is designed to quantitatively measure DNA and RNA oxidized guanosine species. The assay detects all three oxidized guanine species; 8-hydroxy-2’-deoxyguanosine from DNA, 8-hydroxyguanosine from RNA and 8-hydroxyguanine from DNA or RNA. These species may be present in serum, plasma, saliva, urine, dried fecal samples, and tissue culture media samples.

An 8-hydroxy-2’-deoxyguanosine (8-OHdG) stock solution is provided to generate a standard curve for the assay and all samples should be read off the standard curve. Standards or diluted samples are pipetted into a clear microtiter plate coated with an antibody to capture rabbit antibodies. An 8-hydroxyguanosine conjugate is added to the standards and samples in the wells. The binding reaction is initiated by the addition of a peroxidase-labeled mouse monoclonal antibody to 8-hydroxy-2’-deoxyguanosine to each well. After a 2 hour incubation the plate is washed and substrate added. The substrate reacts with the peroxidase labeled antibody that has reacted with the bound conjugate. After a short incubation, the reaction is stopped and the intensity of the generated color is detected in a microtiter plate reader capable of measuring 450nm wavelength. The concentration of the 8-hydroxy-2’-deoxyguanosine in the sample is calculated, after making suitable correction for the dilution of the sample, using software available with most plate readers.