EpiQuik 8-OHdG DNA Damage Quantification Direct Kit (Colorimetric)

Product Description

• Easy-to-follow steps for convenience and speed
• High sensitivity (see Fig. 2) and High specificity (see Fig. 3)
• Direct detection of 8-OHdG using intact DNA
• Detection accuracy is highly correlated with 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 DNA Damage Quantification Direct Kit (Colorimetric) is a complete set of optimized buffers and reagents to colorimetrically detect and quantify oxidative DNA damage (8-OHdG) directly using DNA isolated from any species such as mammals, plants, 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 & Procedure

The EpiQuik™ 8-OHdG DNA Damage Quantification Direct Kit (Colorimetric) contains all reagents necessary for the quantification of Oxidative DNA damage (8-OHdG). In this assay, DNA is bound to strip wells that are specifically treated to have a high DNA affinity. 8-OHdG is detected using capture and detection antibodies. The detected signal is enhanced and then quantified colorimetrically by reading the absorbance in a microplate spectrophotometer. The amount of 8-OHdG is proportional to the OD intensity measured.

 Starting Materials & Input Amount

DNA amount can range from 100 ng to 300 ng per reaction. An optimal amount is 300 ng per reaction. Starting DNA may be in water or in a buffer such as TE.

 Safe and Convenient

All the necessary reagents, including negative controls and positive controls, for the quantification of 8-OHdG are conveniently packaged in the kit. The direct colorimetric quantification of DNA samples replaces obsolete or inferior methods and eliminates the need for DNA digestion/denaturation, radioactivity, extraction, or chromatography.

 Responsive, Reliable, and Practical

Based on its working principle and the microplate format, the kit can be practically and routinely used for any species in a variety of forms including cultured cells, fresh and frozen tissues, and paraffin-embedded tissues. To demonstrate the capabilities of the kit, it has been successfully used for quantifying the content of 8-OHdG in DNA from human kidney, liver, and mouse brain tissues. The percentage of 8-OHdG measured by the kit is similar and comparable to that detected by HPLC methods (see Table 1 and Fig.3).

Table 1. Comparison of Specifications with competitors

Fig.1. 8-OHdG standard control was added into the assay wells at different concentrations and then measured with the EpiQuik™ 8-OHdG DNA Damage Quantification Direct Kit.

Fig.2. Oligos containing 8-OHdG, or 8-OHG or only dG were added into the assay wells at the different concentrations and then measured with the EpiQuik 8-OHdG DNA Damage Quantification Direct Kit (Colorimetric).

Fig.3. Percentage of 8-OHdG in different tissues measured with the EpiQuik 8-OHdG DNA Damage Quantification Direct Kit.