EpiQuik 8-OHdG DNA Damage Quantification Direct Kit

EpiQuik 8-OHdG DNA 損傷標記定量檢測套組

8-hydroxy-2’-deoxyguanosine (8-OHdG or 8-oxo-dG) is an oxidized derivative of deoxyguanosine and is generated by hydroxyl radicals, singlet oxygen, and one-electron oxidants in cellular DNA. As a modified nucleoside base, 8-OHdG is considered important not only because of its abundance but also because of its mutagenic potential through G-to-T transversion mutations upon replication of DNA. 8-OHdG also participates in epigenetic regulation of gene activation/repression by inhibiting the binding affinity of MBD protein to the CpG sites of DNA. Currently, 8-OHdG is widely accepted as a sensitive marker of oxidative DNA damage and oxidative stress. Evidence shows that increased levels of 8-OHdG are closely correlated with exposure to harmful environmental factors such as ionizing radiation, industrial chemicals, air pollution, cigarette smoking, and cancer chemotherapy.

Several chromatography-based techniques such as HPLC-ED and LC-MS are used for detecting 8-OHdG in tissues and cells. However these methods are time consuming and have low throughput with high costs. The currently used competitive ELISA methods are also not conveniently applicable for cell/tissue 8-OHdG detection because they are less accurate and have an inability to use intact DNA isolated from cells or tissues directly.

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.

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實驗原理

The EpiQuik™ 8-OHdG DNA Damage Quantification Direct Kit 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.

(Colorimetric) 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.

(Fluorometric) 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.

Schematic procedure for the EpiQuik™ 8-OHdG DNA Damage Quantification Direct Kit (Colorimetric).

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.

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).

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

Comparison of the EpiQuik™ 8-OHdG DNA Damage Quantification Direct Kit and HPLC-based methods for the quantification of 8-OHdG, in two DNA sample types.

產品列表

貨號	產品	數量
P-6003-48	EpiQuik 8-OHdG DNA Damage Quantification Direct Kit (Colorimetric) 8-OHdG DNA損傷定量分析試劑套組(比色法) (組) 廠牌:Epigentek
P-6003-96	EpiQuik 8-OHdG DNA Damage Quantification Direct Kit (Colorimetric) 8-OHdG DNA損傷定量分析試劑套組(比色法) (組) 廠牌:Epigentek
P-6004-48	EpiQuik 8-OHdG DNA Damage Quantification Direct Kit (Fluorometric) 8-OHdG DNA損傷定量分析試劑套組(螢光法) (組) 廠牌:Epigentek
P-6004-96	EpiQuik 8-OHdG DNA Damage Quantification Direct Kit (Fluorometric) 8-OHdG DNA損傷定量分析試劑套組(螢光法) (組) 廠牌:Epigentek

EpiQuik™ 8-OHdG DNA Damage Quantification Direct Kit

• easy-to-follow steps for convenience and speed. The entire procedure can be completed within 3 hours and 45 minutes.
• High sensitivity, of which the detection limit can be as low as 2 pg of 8-OHdG
• High specificity by detecting only 8-OHdG without cross-reactivity to 8-OHdG analogues
• Direct detection of 8-OHdG using intact DNA, which eliminates interference from high molecular weight compounds, such as carbohydrates and proteins that are often seen in competitive 8-OHdG assays.
• Detection accuracy is highly correlated with and close to HPLC or LC-MS analysis.
• Highly convenient assay with direct use of DNA isolated from cells or tissues, without the need for DNA digestion or hydrolysis.
• Universal positive and negative controls are included, which are suitable for quantifying 8-OHdG from any species.
• Simple, reliable, and consistent assay conditions.