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The multispecies 8-Hydroxydeoxyguanosine (8-OHdG) ELISA quantitates 8-OHdG in serum, plasma, and other biological fluids
Principle of the method:
The Competitive multispecies ELISA research-use-only kit is designed to quantitatively measure 8-Hydroxydeoxyguanosine (8-OHdG) independent of species. A 8-OHdG standard is provided to generate a standard curve for the assay and all samples are read off a user-generated standard curve. Standards or diluted samples are pipetted into a coated microtiter plate and Biotinylated Detection Ab specific to 8-OHdG is added. Excess conjugate and unbound sample or standard are washed from the plate, and Avidin conjugated to Horseradish Peroxidase (HRP) is added to each well and incubated. Then a TMB substrate solution is added to each well. The enzyme-substrate reaction is terminated by the addition of stop solution and the color change is measured on a microplate reader. The concentration of 8-OHdG in the samples is then determined by comparing the OD of the samples to the standard curve.
Rigorous validation:
Each manufactured lot of this ELISA kit is quality tested for criteria such as sensitivity, specificity, precision, and lot-to-lot consistency. See manual for more information on validation.
Oxygen is necessary for aerobic life, it can also participate in potentially toxic reactions involving oxygen free radicals and transition metals such as iron that damage membranes, proteins, and nucleic acids. n a normal human cell, there is a steady accumulation of DNA lesions with time. Substantial parts of these lesions are due to endogenous factors that damage DNA. These include reactive oxygens pecies derived from oxidative respiration. Additionally, malignant cells can produce hydrogen peroxide at levels as high as those characteristics for stimulated polymorphonuclear leukocytes. Some tumors may stimulate the defense systems of the body so that they react against the tumor to produce cytokines. Some cytokines can produce large amounts of ROS. 8-OHdG is one of the most widely studied lesions of this type. The presenceof 8-OHdG residues in DNA leads to GC to TA transversion unless repairs are made before DNA replication. Therefore, the presence of 8-OHdG may lead to mutagenesis. Furthermore, many observations indicate a direct correlation between 8-OHdG formation and carcinogenesis in vivo. It is generally accepted that the repair products of cellular DNA, for example 8-OHdG, are excreted into the urine without further metabolism. Therefore, 8-OHdG has been proposed as a urinary biomarker for in vivo oxidative DNA lesions.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
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