Urinary 8-Hydroxydeoxyguanosine (8-OHdG) ELISA Kit

Advantages of the NWLSS™ Urinary 8-OHDG ELISA:

• Utilizes the N45.1 clone for highest possible specificity for the oxidative DNA damage product 8-OHDG.
• Does not crossreact with products of RNA oxidation such as 8-hydroxy-guanine (8-OHGua) and 8-hydroxy-guanosine (8-OHG).
• Measures 8-OHDG in urine without the need for tedious, time-consuming extractions.

Introduction

Oxidative stress is known to play an important role in the development of various diseases and aging process. Under conditions of oxidative stress, 8-hydroxy-2'-deoxyguanosine (8-OHdG) is formed when DNA is oxidatively damaged by reactive oxygen species (ROS). 8-OHdG is one of the most sensitive biomarkers for oxidative stress and can be detected in urine, plasma and/or DNA isolated from cells and tissues in humans and animals.

Test Principle

The NWLSS™ 8-OHdG ELISA assay uses a competitive format wherein a murine monoclonal antibody to 8-OHdG (Primary Antibody) and sample or standard are added to a microtiter plate which has been precoated with 8-OHdG. Sample or calibrator 8-OHdG competes with plate-bound 8-OHdG for binding with the antibody. Accordingly, higher concentrations of sample or calibrator lead to reduced binding of the antibody to the 8OHdG coated on the plate. A subsequent wash step removes any free 8-OHdG/antibody adduct leaving stationary plate bound 8-OHdG complexed to antibody for later detection. Anti-murine antibody conjugated to horse radish peroxidase (HRP-Conjugate) is then added to the plate. HRP-conjugate binds to remaining murine anti-8-OHdG and unbound HRP-conjugate is removed in another wash step. Addition of 3,3',5,5'tetramethylbenzidine (TMB Substrate) results in blue color development proportional to the amount of anti 8-OHdG antibody bound to the plate and inversely proportional to theconcentration 8-OHdG in original samples or calibrators applied to the plate. The reaction is terminated by addition of phosphoric acid (Stop Solution) producing yellow color with measurable absorbance at 450 nm.