Urinary 8-Hydroxydeoxyguanosine (8-OHdG) ELISA Kit

NWLSS | Northwest Life Science Specialties, LLC.
Urinary 8-Hydroxydeoxyguanosine (8-OHdG) ELISA Kit
NWK-8OHDG01 $945.00 each

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The NWLSS™ Urinary 8-OHDG ELISA is intended for quantitative detection of the oxidized DNA adduct 8-OHdG in urine. For samples expected to contain lower amounts of 8-OHdG such as plasma and/or DNA extracts isolated from cell lysates or tissue homogenates, we recommend the High Sensitivity 8-Hydroxydeoxyguanosine (8-OHdG) ELISA [NWK-8OHDG02]

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.

We would like to measure Catalase activity in tissue samples. Can we prepare the tissue homogenate samples using 0.05M PBS, pH7.0?

Normal 1X PBS is typically 10 mM so the .05M PBS you are thinking of using may be at 5X strength. We cannot say for sure whether the increased strength would be an issue with the CAT01 assay so would simply recommend diluting the solution to 10 mM PBS for homogenizing your samples.
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For Research Use Only

Catalog Number: NWK-8OHDG01
Format: 96 wells presented as 1 plates sectionable as 8 well individual strips
Sample Requirements: Recommended for Urine. Plasma & DNA Extracts from cells or tissue are best assayed with product [NWK-8OHDG02].
Sensitivity: 0.5 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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