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- Assay Type Competitive ELISA
- Sample Types Serum, Plasma, Saliva, Urine, Fecal Extracts, Digested DNA, Tissue Culture Media
- Sensitivity 50.9 pg/mL
- Species Mammals
- Assay Duration 2.5 Hours
- Samples/Plate 38 in Duplicate
- Readout Colorimetric, 450 nm
- Standard Curve
The DNA Damage ELISA Kit quantitatively measures DNA and RNA oxidized guanosine species in serum, plasma, saliva, urine, fecal extracts, digested DNA, and tissue culture media. The DNA Damage ELISA Kit is a competitive ELISA with a run time of 2.5 hours. Please read the complete kit insert for more information before performing this assay.
Use our provided 8-Hydroxy-2’-deoxyguanosine (8-OHdG) standard to generate a standard curve for the assay. Pipette the standards or diluted samples into a transparent microtiter plate coated with our goat anti-mouse IgG antibody. Add the 8-hydroxyguanosine conjugate and the peroxidase-labeled mouse monoclonal antibody. Then incubate the mixture covered at room temperature, shaking for 2 hours. The immunological reaction occurs between the anti-8-OHdG antibody, the 8-OHdG antigen in the sample or standard, and the 8-OHdG conjugate. As the 8-OHdG concentration in the sample increases, the bound 8-OHdG-peroxidase conjugate decreases, causing a decrease in signal and vice versa.
After the 2-hour incubation, wash away the excess 8-OHdG-peroxidase conjugate and add the TMB substrate. The TMB substrate reacts with the bound 8-OHdG-peroxidase conjugate generating a signal detected by a plate reader at 450nm. Use the intensity and the standard curve to calculate the 8-OHdG concentration in the samples.
In vivo biological processes generate free radicals and other reactive species which cause oxidative damage to biomolecules. Multiple antioxidant repair systems and the replacement of damaged nucleic acids, proteins, and lipids combat this oxidative damage. The generation of intracellular free radical species (ROS) occurs during normal metabolism. Ultraviolet radiation or ionizing radiation generates extracellular forms of ROS. A cellular function may be stopped if DNA damage corrupts the information contained within the genome.
Studies show that continuous oxidative damage to DNA significantly contributes to the age-related development of major cancers, such as colon, breast, rectum, and prostate. 8-hydroxy-2’-deoxyguanosine (8-OHdG) is a major product of DNA oxidation. 8-OHdG is a marker of oxidative stress and carcinogenesis. 8-OHdG is physiologically formed during the repair of damaged DNA in vivo by exonucleases. The resulting 8-OHdG is excreted without further metabolism into the urine.