We offer a suite of DNA/RNA Damage research products to help determine the relationship between oxidative damage and disease.
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Introduction to Oxidative Damage
Oxidative stress damage plays an important role in the etiology and progression of a multitude of human pathologies, including oncologic, cardiovascular, and autoimmune diseases and disorders. Through reactive oxygen species (ROS), oxidative stress can result in cellular damage, enzyme inactivation, and epigenetic impairment of DNA. A better understanding of oxidative stress damage—disease relationships can therefore be beneficial for diagnostic and therapeutic purposes.
As a consequence, various biomarkers have been identified for oxidative stress damage, with the most well-studied being: oxidized nucleoside derivatives like 8-hydroxy-2’-deoxyguanosine (8-OH-dG) and 8-hydroxyguanosine (8-OH-G) that are generated by ROS in cellular DNA and RNA, respectively; the ROS hydrogen peroxide (H2O2) and its catalyzing enzyme superoxide dismutase (SOD); and histone H2AX phosphorylation, particularly at serine 139, which manifests during the DNA damage response.
Most available methods used to investigate these biomarkers are time-consuming and have low throughput, low accuracy, and high costs. By leveraging its EpiQuik™ technologies, EpiGentek has developed a series of innovative research tools to address those issues. Our comprehensive product suite for oxidative stress damage assessment includes the following features and advantages:
Fast procedure
High sensitivity and specificity
Simple, reliable, and consistent assay conditions
Colorimetric/fluorometric ELISA-based assays with easy-to-follow steps for convenience and speed
Assay flexibility via strip-well microplate format for either manual or high throughput analysis
8-OH-dG and 8-OH-G
8-hydroxy-2′-deoxyguanosine (8-OH-dG) and 8-hydroxyguanosine (8-OH-G) are oxidized guanosine derivatives generated in cellular DNA and RNA, respectively, by hydroxyl radicals, singlet oxygen, and one-electron oxidants. 8-OH-dG and 8-OH-G are considered important not only due to their abundance but also because of their mutagenicity through mismatched pairings that can cause transversions and induce errors in gene expression. These modified nucleoside bases also participate in epigenetic regulation of gene activation and repression by affecting the recruitment of structural proteins and subsequent chromatin modification.
While oxidatively damaged DNA can be quickly repaired via base excision repair pathways intracellularly, oxidized RNA can remain in cells for hours after a short insult of oxidative stress, and this oxidative RNA mark could be identified as an early indicator of cell death. Evidence shows that increased levels of 8-OH-dG and 8-OH-G are closely correlated with exposure to harmful environmental factors such as ionizing radiation, industrial chemicals, air pollution, cigarette smoking, and cancer chemotherapy. It has also been demonstrated that increased concentrations of 8-OH-dG and 8-OH-G are pathogenically linked to a variety of age-associated diseases, including cancer, coronary heart disease, diabetes, and neurodegenerative diseases such as Alzheimer's disease.
Hydrogen Peroxide
Hydrogen peroxide (H2O2) is a reactive oxygen species generated in cellular oxygen metabolism. Abnormal H2O2 levels contribute to oxidative cell damage and the progression of diseases such as inflammation, ischemia, and neurodegeneration. H2O2 is a major marker of ROS release from cells or organelles such as mitochondria. It can also be an indicator of oxidase activity as a co-product of oxidase-catalyzed reactions, such as those facilitated by NADPH and glucose oxidases. Therefore, it is important to detect H2O2 from various biological samples in determining how oxidative stress modulates varied intracellular pathways under physiological and pathological conditions.
Superoxide Dismutase
Superoxide dismutase (SOD) is one of the most important enzymes in defending against oxidative stress. SOD catalyzes the dismutation of superoxide anion (O2- ) into H2O2 and molecular oxygen (O2), serving as a key antioxidant, but can also inflict many types of cell injury if dysregulated. Determination of SOD activity is thus necessary in various research fields related to human diseases.
Histone H2AX Phosphorylation
Studies have shown that in vitro ROS exposure results in high frequencies of DNA single-strand breaks (SSBs) and double-strand breaks (DSBs). DSBs caused by cellular exposure to genotoxic agents or produced by inherent metabolic processes initiate a rapid and highly coordinated series of molecular events resulting in DNA damage signaling and repair. There is increasing evidence for the role of chromatin in DNA damage responses, and histone modifications and chromatin modulation have been implicated in genome stability. Phosphorylation of histone H2AX at serine 139 to form γH2AX is one of the earliest chromatin modification events during the DNA damage response and is important for the coordination of signaling and repair activities. Therefore, phospho-H2AX (Ser139) can be used as a rapid and sensitive marker for DNA damage and apoptosis.
Sample Preparation
Proper isolation of DNA, RNA, and proteins is an important precursor to having a successful experiment. EpiGentek offers a variety of sample preparation kits that are suitable for nearly any workflow. Choose the right product for your research needs.
For isolation of small fragment circulating ccfDNA from plasma or serum
8-OH-dG DNA Damage Quantification Kits
8-hydroxy-2’-deoxyguanosine (8-OHdG) is an important biomarker of oxidative DNA damage associated with mutagenesis and carcinogenesis. Many methods for detecting 8-OHdG in DNA samples are time-consuming, costly and lack the ability to use intact DNA from cells or tissues. EpiGentek’s ELISA-like 8-OHdG DNA damage quantification kits can detect and quantify oxidative DNA damage (8-OHdG) directly from DNA in an easy-to-follow procedure that takes less than 4 hours.
Fluorometric assay for quantitating 8-OH-dG in DNA samples (LOD = 1 pg of 8-OH-dG)
8-OH-G RNA Damage Quantification Kit
8-hydroxyguanosine (8-OhG or 8-oxoG) is an abundant form of oxidative RNA damage. Evidence has shown that mutagenic mispairing from 8-OhG can produce errors in gene expression. Plus, this modified nucleoside base has been linked to a variety of age-associated diseases. To assist the researcher in studying the importance of 8-OhG, EpiGentek offers an ELISA-like kit that specifically detects oxidative RNA damage (8-OhG) status in using RNA isolated from any species in just 3 hours.
Accurately measure hydrogen peroxide(H2O2) from different samples either in-situ (living cells) or ex-situ (cell/tissue extracts and body fluids) using our EpiQuik In-Situ and Ex-Situ Hydrogen Peroxide (H2O2) Assay Kit. The kit contains everything needed to detect H202 from a sample and takes just 10 minutes for results.
Fast and sensitive measurement of H2O2 from various samples, including living cells, cell/tissue extracts, and body fluids
SOD Enzyme Activity/Inhibition Assay
Superoxide dismutase (SOD) is one of the most important enzymes in defending oxidative stress. There are several methods used for measuring SOD activity; however, most of these methods have some drawbacks in selectivity, rapidity, and convenience. The EpiQuik Superoxide Dismutase Activity/Inhibition Assay Kit addresses these problems by using a unique procedure to measure SOD activity.
Detects SOD activity in cell/tissue extracts by measuring color production of a reaction mediated by the SOD substrate superoxide anion
Phospho-H2AX (Ser139) Assay Kit
Detect DNA damage or apoptosis by measuring phosphorylation of H2AXSer139 in situ using our EpiQuik In Situ DNA Damage Assay Kit. The kit is ready to use and includes all the essential components necessary for specifically measuring DNA damage in situ via phospho H2AXSer139 detection.
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