Overview

  • Product name8-hydroxy 2 deoxyguanosine ELISA Kit
  • Assay typeSandwich (quantitative)
  • Sensitivity
    30 pg/ml
  • Range
    30 pg/ml - 100 pg/ml
  • 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 applicationsELISAmore details

Properties

  • 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.
  • Alternative names
    • 8 OH dG

Applications

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.

Application Abreviews Notes
ELISA Use at an assay dependent dilution.

8-hydroxy 2 deoxyguanosine ELISA Kit images

  • 8OH2dG measured in biological fluids, background signal subtracted (duplicates +/- SD).
  • ab101245 used in ELISA. Homogenized murine colon tissue sections 5µg DNA.

    See Abreview

  • Typical standard curve

    50% B/B0 - 115 pg/ml
    Detection Limit (80% B/B0) - 33 pg/ml

  • 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

Protocols

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) »

See 1 Publication for this product

Product Wall



Here is the response from my colleagues in the lab:

You can use whatever nuclease P1 to digest the DNA that you want. We mention the specific one in the kit booklet, because that is the particular one that we used for the kit. Howev...

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Gracias por tu respuesta.

Tan pronto como tengamos nuevas referencias de publicaciones para este kit, actualizaremos su datasheet.

En cualquier caso, cualquier información que recibamos por parte de los usuarios de nuestros p...

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Gracias por contactarnos.



Todas las referencias y usos de este kit disponibles están detalladas en la datasheet. No tenemos constancia de su uso en bacterias. Sin embargo, la fuente de la que proceda 8OHdG no es dependiente d...

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Ihave issued a freekit, 8-hydroxy 2 deoxyguanosine ELISA Kit, ab101245,with the order number 1070428.

To check the status of the order please contact our Customer Service team and reference this number.

Again, I apologize for any ...

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Thank you for contacting us.

Theoretically the kit will be suitable for samples irrespective of the species.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Thank you for contacting Abcam.
I have talked to the lab and the antibody in the kit ab101245, is actually one that we have in our catalogue.
The antibody in ab101245 is actually ab62623, but it's slightly varied in its buffers. The kit produ...

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If stored as directed, the kit can be stored for at least 1 year.

I hope this is helpful. Please contact me again if you have any further questions.

Abreviews

Sample: Mouse Tissue sections (Homogenized colon sections)
Specification: Homogenized colon sections
Amount of Sample: 5 ug DNA µg
Type of Kit Used: ELISA
Positive control: None
Did you use the Abcam Kit protocol?: Yes
Username

Mr. Ahmed Nayef

Verified customer

Submitted Feb 10 2012

Thank you for contacting us. The mass of DNA used is not as important as the amount of 8-hydroxy 2 deoxyguanosine present. This can vary by several orders of magnitude depending on your samples. We typically recommend starting with around 50ug...

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Thanks for your call today and for your patience while I have been in touch with lab. Since P1 cuts without preference and DNase 1 is preferential for cuts adjacent to pyrimidine, and my contact at the lab recommends using P1 if possible. Other meth...

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"