High Sensitivity 8-Hydroxydeoxyguanosine (8-OHdG) ELISA Assay Kit

NWLSS | Northwest Life Science Specialties, LLC.
nwk-8ohdg02
NWK-8OHDG02 $945.00 each

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The NWLSS™ High Sensitivity 8-OHDG ELISA is intended for quantitative detection of the 8-OHdG adduct in plasma, and/or DNA extracts isolated from cell lysates or tissue homogenates. This assay has better sensitivity than the (NWK-8OHDG01)but requires overnight incubation. When testing urine samples, same day results are possible when using our Urinary 8-Hydroxydeoxyguanosine (8-OHdG) ELISA Kit.

Advantages of the NWLSS™ High Sensitivity 8-OHDG ELISA:

  • Utilizes the N45.1 clone for highest possible specificity for the oxidative DNA product 8OHDG.
  • Does not crossreact with products of RNA oxidation such as 8-hydroxy-guanine (8-OHGua) and 8-hydroxy-guanosine (8-OHG).
  • Higher sensitivity allows quantification of 8-OHDG down to 125 pg/mL

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. The figure below depicts the oxidation 2'-deoxyguanosine (2'dG) to form the 8-OHdG biomarker.

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. During an overnight incubation, 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 8-OHdG 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 8OHdG antibody bound to the plate and inversely proportional to the concentration 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.

For Research Use Only

Catalog Number: NWK-8OHDG02
Format: 96 wells presented as 1 plates sectionable as 8 well individual strips
Sample Requirements: Recommended for Plasma & DNA extracts from cells or tissue
Sensitivity: 0.125 ng/mL
Storage and Stability: 9 months from date of manufacture when stored at specified temperature
Kit Contents: Precoted Microwell strips and strip frames
8-OHdG Standards
Dilution Buffer
Primary Antibody
Primary Antibody Buffer
Secondary Antibody
Secondary Antibody Buffer
TMB Substrate
Wash Buffer
Stop Solution
Plate Seals

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