Description
Key facts
Detection method:Colorimetric,
Sample types:Saliva, Urine, Plasma, Tissue Extracts, Serum, Cell Lysate,
Assay type:Competitive,
Sensitivity:= 0.59 ng/mL,
Range:0.94 - 60 ng/mL,
Assay time:2h,
Assay Platform:Pre-coated microplate (12 x 8 well strips)
Product details:
8-hydroxy 2 deoxyguanosine ELISA Kit (8-OHdG) ab201734 is a competitive ELISA kit to measure 8-Hydroxy-2'-deoxyguanosine with a sensitivity of 0.59 ng/mL.
How the assay works
8-hydroxy 2 deoxyguanosine ELISA Kit ab2017341 is a competitive ELISA. There are 2 types of competitive ELISAs:
antigen down
: The sample antigen competes with a reference antigen for binding to a specific amount of labeled antibody. The reference antigen is pre-coated on a multi-well plate. The sample is added to the plate wells with a labeled antibody. Depending on the amount of antigen in the sample, more or fewer free antibodies will be available to bind the reference antigen. This means that the more of the antigen there is in the sample, the less reference antigen will be detected and the weaker the signal will be.
antibody down
: the labeled antigen and the sample antigen (unlabeled) compete for binding to the primary antibody, which is immobilized on the plate. The lower the amount of antigen in the sample, the stronger the signal due to more labelled antigen in the well.
Assay specificity
Our ELISA kits are rigorously validated to ensure the highest level of consistency and reproducibility.
8-hydroxy 2 deoxyguanosine ELISA Kit (8-OHdG) ab201734 protocol summary
- Add standards or samples to the wells, incubate 2 hrs
- Wash, then add 50 µl of Biotinylated Antibody per well. Incubate 1 hr
- Wash, then add 50 µl of SP Conjugate per well. Incubate 30 min
- Wash, then add 50 µl of Chromogen Substrate per well. Incubate 20 min
- Add 50 µl of Stop Solution per well. Read at 450 nm immediately
It is important to note that the 8-OHdG antibody used in this assay recognizes both free 8-OHdG and DNA-incorporated 8-OHdG. Since complex samples such as plasma, cell lysates, and tissues are comprised of mixtures of DNA fragments and free 8-OHdG, concentrations of 8-OHdG 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.
8-hydroxy-2-deoxy Guanosine (8-OHdG) is produced by the oxidative damage of DNA by reactive oxygen and nitrogen species and serves as an established marker of oxidative stress. Hydroxylation of guanosine occurs in response to both normal metabolic processes and a variety of environmental factors (i.e., anything that increases reactive oxygen and nitrogen species). Increased levels of 8-OHdG are associated with the aging process as well as with a number of pathological conditions including cancer, diabetes, and hypertension.
In complex samples, such as plasma, cell lysates, and tissues, 8-OHdG can exist as either the free nucleoside or incorporated in DNA. Once the blood enters the kidney, free 8-OHdG is readily filtered into the urine, while larger DNA fragments remain in the bloodstream. Because of the complexity of plasma samples, urine is a more suitable matrix for the measurement of free 8-OHdG than plasma. Urinary levels of 8-OHdG range between 2.7- 13 ng/mg creatine, while plasma levels of free 8-OHdG have been reported to be between 4-21 pg/mL as determined by LC-MS.
Properties and Storage Information:
Shipped at conditions-Blue Ice, Appropriate short-term storage conditions--20°C, Appropriate long-term storage conditions-Multi, Storage information-Please refer to protocols
Supplementary Information:
This supplementary information is collated from multiple sources and compiled automatically.
8-Hydroxy-2'-deoxyguanosine (8-OHdG) also known as 8-hydroxyguanine or simply 8-OHdG is a well-known marker of oxidative DNA damage. This compound forms through the oxidative modification of deoxyguanosine a nucleoside component of DNA mainly by reactive oxygen species (ROS). 8-OHdG is a stable product with a molecular mass of 283.26 g/mol. It is found in various tissues and is excreted in urine which makes it a useful non-invasive biomarker for assessing oxidative stress levels in the body.
Biological function summary
8-hydroxy-2-deoxyguanosine plays a significant role in indicating cellular oxidative damage but it doesn't act as a signaling molecule or enzyme. Instead it serves as a biomarker that results from the damage. Oxidative stress caused by reactive oxygen species can lead to the formation of 8-OHdG and indicates potential mutations that may arise if the damage goes unrepaired. Its formation does not involve complex protein assemblies but results from the attack on DNA by ROS highlighting cellular environments exposed to oxidative conditions.
Pathways
Several factors cause oxidative stress leading to the generation of 8-OHdG linking it to key biological pathways such as the DNA damage response and repair pathways. The base excision repair (BER) pathway is important for recognizing and repairing the damage introduced by 8-OHdG. Proteins such as 8-oxoguanine glycosylase (OGG1) directly interact with 8-OHdG during the repair process excising the damaged base from DNA to prevent mutations and maintain genomic integrity.
Researchers associate elevated levels of 8-OHdG with various conditions linked to oxidative stress such as cancer and neurodegenerative diseases like Alzheimer's disease. In cancer the accumulation of 8-OHdG can provide evidence of increased oxidative DNA damage which may lead to mutations and carcinogenesis. Alzheimer’s disease patients often exhibit elevated 8-OHdG levels indicating oxidative damage in neural tissues. The protein OGG1 plays a pivotal role in relationship with these diseases serving as an important enzyme for repairing the 8-OHdG lesions and potentially impacting disease progression.
