Aldehyde reactive probe (ARP) is used to detect abasic sites (AP sites, depurine/depyrimidine sites) in DNA because of its aldehyde-specific reactivity. ARP reacts with aldehyde and ketone groups, and it adds biotin to AP sites in DNA. The biotin-tagged DNA is then detected using peroxidase-labeled avidin and oxidative chromogenic dyes. The ARP method has been used to detect less than 1 AP site in 1×104 nucleotides of DNA. Dojindo offers a DNA Damage Quantification Kit (Product Code: DK02) containing all the reagents and components necessary to determine 1 to 40 ARP sites per 1×105 base pairs. ARP is highly soluble in water and the stock solution can be stored at 4ºC for one year without significant loss of reactivity.
Labeling Procedure for DNA Abasic Site
1. Prepare 10 mM ARP with water.
2. Adjust the DNA concentration to 100 μg per ml with TE buffer (pH 7.4).
3. Mix equal volumes of the DNA and ARP solutions and incubate at 37ºC for 1 hour.
4. Isolate the ARP-labeled DNA using either ethanol precipitation or membrane filtration tube.
5. Dissolve the DNA pellet in TE to prepare 10-100 μg per ml solution.
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