Journal: Chembiochem : a European journal of chemical biology
Article Title: In Vitro Fluorogenic Real-time Assay of the Repair of Oxidative DNA Damage
Figure Lengend Snippet: A and B) PAGE analysis of 4 μM probe OGR1 radiolabeled at the 5′ end with [ 32 P]-phosphate and reacted with 658 nM hOGG1 (A) or Fpg (B) at 37 °C for various times. Piperidine (200 mM, 95 °C, 5 min), which cleaves the DNA backbone at abasic sites, was added to samples as indicated to identify the 8-oxoguanine excision product and the backbone cleavage product. C and D) MALDI mass spectrometry analysis of probe OGR1 following overnight reaction with hOGG1 (C) or Fpg (D).
Article Snippet: Fpg, hOGG1, Fpg reaction buffer (NEB buffer 1: 10 mM Bis-Tris-Propane-HCl, 10 mM MgCl2 , 1 mM dithiothreitol, pH 7.0 at 25 °C), hOGG1 reaction buffer (NEB buffer 4: 50 mM potassium acetate, 20 mM Tris-acetate, 10 mM magnesium acetate, 1 mM dithiothreitol, pH 7.9 at 25 °C), and BSA (10 mg/mL) were purchased from New England Biolabs.
Techniques: Polyacrylamide Gel Electrophoresis, Mass Spectrometry