Research area: Oxidative stress
DNA / RNA Damage and Repair Assays
cultured cells, fresh and frozen tissues, paraffin-embedded tissues, body fluid samples, etc.
any species such as mammals, plants, fungi, bacteria, and viruses
Detection method: colorimetric
- Easy-to-follow steps for convenience and speed
- High sensitivity (see Fig. 2) and high specificity
- Direct detection of 8-OHdG using intact DNA
- Detection accuracy is highly correlated and close to HPLC or LC-MS analysis
- Direct use of DNA isolated from cells or tissues
- Universal positive and negative controls included
- Suitable for manual or high-throughput analysis
The EpiQuik ™ 8-OHdG Direct DNA Damage Quantification Kit (Colorimetric) is a complete set of buffers and reagents optimized to detect and colorimetrically quantify oxidative DNA damage (8-OHdG) directly using DNA isolated from any species, like mammals, plants, and fungi. , bacteria, and viruses in a variety of forms including, but not limited to, cultured cells, fresh and frozen tissues, paraffin-embedded tissues, and body fluid samples.
Principle and procedure
The EpiQuik ™ 8-OHdG Direct DNA Damage Quantification Kit (Colorimetric) contains all the reagents necessary for the quantification of oxidative DNA damage (8-OHdG). In this assay, DNA is bound to wells that are specifically treated to have a high affinity for DNA. 8-OHdG is detected by capture and detection antibodies. The detected signal is enhanced and then quantified colorimetrically by reading the absorbance on a microplate spectrophotometer. The amount of 8-OHdG is proportional to the DO intensity measured.
Starting materials and input quantity
The amount of DNA can range from 100 ng to 300 ng per reaction. An optimal amount is 300 ng per reaction. The starting DNA can be in water or in a buffer such as TE.
Safe and convenient
All necessary reagents, including negative controls and positive controls, for 8-OHdG quantitation, are conveniently packaged in the kit. Direct colorimetric quantification of DNA samples replaces outdated or inferior methods and eliminates the need for DNA digestion/denaturation, radioactivity, extraction, or chromatography.
Responsive, reliable, and practical
Based on its principle of operation and microplate format, the kit can be practically and routinely used for any species in a variety of ways including cultured cells, fresh and frozen tissues, and paraffin-embedded tissues. To demonstrate the capabilities of the kit, it has been used successfully to quantify the content of 8-OHdG in the DNA of human-mouse kidney, liver, and brain tissues.