8-HYDROXYGUANOSINE; GTX-200UL

Analysis Note

ControlSkin, staining is limited to the nucleus.

Application

This Anti-8-Hydroxydeoxyguanosine Antibody is validated for use in ELISA, IH(P) for the detection of 8-Hydroxydeoxyguanosine.

Immunohistochemistry: >1:200 on paraffin embedded, formalin fixed human brain. It is suggested that the tissues are pretreated with 10 µg/mL of proteinase K for 40 minutes at 37°C prior to staining.

Dewaxed sections treated with 3%H2O2 in methanol for 30 minutes and blocking was with 10% NGS in TBS. Sections were treated with proteinase K 10ug/ml in PBS pH 7.4, 40 minutes 37C prior to blocking. Detection is via DAB. Since the antibody is staining a nucleic acid it is important to use enough proteinase to penetrate the histone barrier.
Optimal working dilutions must be determined by the end user.

General description

8-Hydroxydeoxyguanosine (8OHdG) is a modified base that occurs in DNA due to attack by hydroxyl radicals that are formed as byproducts and intermediates of aerobic metabolism and during oxidative stress. There is increasing evidence to support the involvement of free radical reactions in the damage of biomolecules that eventually lead to several diseases in humans, such as atherosclerosis, cerebral and heart ischemia-reperfusion injury, cancer, rheumatoid arthritis, inflammation, diabetes, aging, and neurodegenerative conditions, such as Alzheimer′s disease.

8OHdG has become increasing popular as a sensitive, stable and integral marker of oxidative damage in cellular DNA. Biomonitoring in humans has demonstrated that 8OHdG can be excreted in the urine, and that a significant increase is caused by exposure to tobacco smoke and ionizing radiation. Because 8OHdG is so well correlated with oxidative stress and damage to DNA, which leads to degenerative disease states, the development of an antibody that can be used to study DNA damage has numerous applications. In addition to the direct study of DNA damage within cells, this antibody has applications in the development of immunoassays that can monitor 8OHdG excretion in the urine and serve as a biomarker of oxidative stress. Industrial uses may extend to the dietary supplement manufacturers, who could benefit from an immunoassay that could be used to test the effectiveness of antioxidants and other nutraceuticals.

CHEMICON′s new goat antiserum to 8-hydroxyguanosine is now available. This antiserum has been shown by ELISA to be completely cross-reactive with 8-hydroxydeoxyguanosine while not cross-reacting with other naturally occurring nucleotides. This antibody should be a valuable tool for scientists working to understand the role of free radical damage in a number of human disease states.

Immunogen

8-Hydroxy-2′-deoxyguanosine and 8-hydroxyguanosine.

Legal Information

CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Specificity

Reacts with 8-Hydroxydeoxyguanosine (8-OHdG) and 8-Hydroxyguanosine (8-OHG). By a competitive ELISA the antibody was 100% inhibited by 8-hydroxydeoxyguanosine and 8-hydroxyguanosine nucleotides. Partial cross-reactivity was observed with 8-mercaptoguanosine (80%) while 8-bromoguanosine showed some cross-reactivity (5%). 2-Deoxyadenosine, 7-methlyguanosine, guanosine monophosphate and guanosine showed no reactivity.