The 8-OHdG ELISA kit is a competitive in vitro enzyme-linked immunosorbent assay for quantitative measurement of
the oxidative DNA adduct 8-hydroxy-2’-deoxyguanosine (8-OHdG) in tissue, urine etc. For research use only. Not for
diagnostic nor medical use. Read entire insert before use.
Kit Contents (Highly Sensitive 8-OHdG Check #NNS-KOG-HS10E-EX)
- 8-OHdG Microtiter plate : precoated with 8-OHdG (812wells, split type)
- Primary antibody : monoclonal antibody specific for 8-OHdG
- Primary antibody solution : phosphate buffered saline
- Secondary antibody : HRP-conjugated antibody
- Secondary antibody solution : phosphate buffered saline
- Chromatic solution : 3,3',5,5'-tetramethylbenzidine
- Diluting solution : hydrogen peroxide/citrate-phosphate buffered saline
- Washing solution(5) : 5 times concentrated phosphate buffered saline**
- Reaction terminating solution: 1M phosphoric acid
- Standard 8-OHdG solution : 0.125, 0.25, 0.5, 1, 4, 10 ng/ml 1vial each
- Plate seal
*All reagent should be refrigerated at 2-8C.
*The kit should not be used beyond 9 months past the manufacturers date stamped on the exterior of the box.
**Dilute Washing solution by 5 times (v/v) with distilled water for use.
Summary of Assay Procedure
- The 8-OHdG monoclonal antibody and the sample or standard are added to the microtiter plate which has been precoated with 8-OHdG. The 8-OHdG monoclonal antibody reacts
competitively with the 8-OHdG bound on the plate and the 8-OHdG in samples solution. Therefore higher concentrations of 8-OHdG in the sample solution lead to a reduced binding of the antibody to the 8-OHdG on the plate.
- The antibodies which are bound to the 8-OHdG in the sample are washed away from the antibodies that have bound to the 8-OHdG coated on the plate.
- An enzyme-labeled secondary antibody, which is added to the plate, binds to the monoclonal antibody which is bound to the 8-OHdG coated on the plate.
- Unbound enzyme-labeled secondary antibody is removed by a wash step.
- Addition of a chromatic substrate results in the development of color in proportion to the amount of antibody bound to the plate.
- The color reaction is terminated and the absorbance is measured.