8-hydroxy 2 deoxyguanosine ELISA Kit (ab101245)
- Product name8-hydroxy 2 deoxyguanosine ELISA Kit
- Assay typeSandwich (quantitative)
- Product overview
ab101245 8-hydroxy 2 deoxyguanosine (8-OH-dG) ELISA Kit is a competitive assay that can be used for the quantification of 8-OH-dG in urine, cell culture, plasma, and other sample matrices. The ELISA Kit utilizes an anti-mouse IgG-coated plate and a tracer consisting of an 8-OH-dG-enzyme conjugate. This format has the advantage of providing low variability and increased sensitivity compared to assays that utilize an antigen-coated plate. Our ELISA Kit typically displays IC50 (50% B/B0) and IC80 (80% B/B0) values of approximately 100 and 30 pg/ml, respectively.
It is important to note that the 8-OH-dG antibody used in this assay recognizes both free 8-OH-dG and DNA-incorporated 8-OH-dG. Since complex samples such as plasma, cell lysates, and tissues are comprised of mixtures of DNA fragments and free 8-OH-dG, concentrations of 8-OH-dG reported by ELISA methodology will not coincide with those reported by LC-MS where the single nucleoside is typically measured. This should be kept in mind when analyzing and interpreting experimental results.
This assay is based on the competition between 8-hydroxy-2-deoxy guanosine (8-OH-dG) and a 8-OH-dG-acetylcholinesterase (AChE) conjugate (8-OH-dG Tracer) for a limited amount of 8-OH-dG Monoclonal Antibody. Because the concentration of the 8-OH-dG Tracer is held constant while the concentration of 8-OH-dG varies, the amount of 8-OH-dG Tracer that is able to bind to the 8-OH-dG Monoclonal Antibody will be inversely proportional to the concentration of 8-OH-dG in the well. This antibody-8-OH-dG complex binds to goat polyclonal anti-mouse IgG that has been previously attached to the well. The plate is washed to remove any unbound reagents and then Ellman’s Reagent (which contains the substrate to AChE) is added to the well. The product of this enzymatic reaction has a distinct yellow color and absorbs strongly at 412 nm. The intensity of this color, determined spectrophotometrically, is proportional to the amount of 8-OH-dG Tracer bound to the well, which is inversely proportional to the amount of free 8-OH-dG present in the well during the incubation.
AChE has several advantages over other enzymes commonly used for enzyme immunoassays. Unlike horseradish peroxidase, AChE does not self-inactivate during turnover. This property of AChE also allows redevelopment of the assay if it is accidentally splashed or spilled. In addition, the enzyme is highly stable under the assay conditions, has a wide pH range (pH 5-10), and is not inhibited by common buffer salts or preservatives. Since AChE is stable during the development step, it is unnecessary to use a ‘stop’ reagent, and the plate may be read whenever it is convenient.
- Tested applicationsELISA more details
- Storage instructionsStore at +4°C. Please refer to protocols.
Components 1 x 96 tests 8 well strips 12 units 8-hydroxy-2-deoxy Guanosine Ache tracer 1 x 100 units 8-hydroxy-2-deoxy Guanosine antibody in PBS 1 x 100 units 8-hydroxy-2-deoxy Guanosine EIA standard 1 unit EIA antiserum dye 1 unit EIA buffer concentrate 2 x 10ml EIA tracer dye 1 unit Ellman’s Reagent 3 x 100 units Tween 20 1 x 3ml Wash Buffer Concentrate 1 x 5ml
- Research Areas
- RelevanceOxidative damage of DNA or RNA by reactive oxygen and nitrogen species produces 8-hydroxy Guanine. This can serve as a measure of oxidative stress.
- 8 OH dG
Our Abpromise guarantee covers the use of ab101245 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
8-hydroxy 2 deoxyguanosine ELISA Kit images
Recovery of 8-hydroxy-2-deoxy Guanosine from urine
Urine samples were spiked with 8-OH-dG, diluted as described in the Sample Preparation section and analyzed using the 8-OH-dG EIA Kit. The y-intercept corresponds to the amount of 8-OH-dG in unspiked urine. Error bars represent standard deviations obtained from multiple dilutions of each sample
Typical standard curve
50% B/B0 - 115 pg/ml
Detection Limit (80% B/B0) - 33 pg/ml
ab101245 used in ELISA. Homogenized murine colon tissue sections 5µg DNA.
References for 8-hydroxy 2 deoxyguanosine ELISA Kit (ab101245)
This product has been referenced in:
- Zhou X et al. Variation in dietary salt intake induces coordinated dynamics of monocyte subsets and monocyte-platelet aggregates in humans: implications in end organ inflammation. PLoS One 8:e60332 (2013). Read more (PubMed: 23593194) »