Thin layer chromatography-based assay of O6-methylguanine-DNA methyltransferase activity in tissue.
Expression of the DNA repair enzyme O(6)-methylguanine-DNA methyltransferase (MGMT) in tumor correlates with resistance to alkylating agents, and depletion of MGMT activity can enhance chemotherapy-induced tumor cytotoxicity. The most common assays of MGMT activity are time-consuming and employ radioactivity. The assay described here uses a fluorescently labeled O(6)-benzylguanine derivative in conjunction with thin layer chromatography to eliminate the use of radioactivity and allows MGMT activity to be rapidly measured in minimally prepared cell or tissue extracts.
Robinson
Cliff
C
Department of Radiation Oncology, Washington University in St. Louis, St. Louis, MO 63110, USA.
Palomo
Juan
J
Vogelbaum
Michael A
MA
D045744:Cell Line, Tumor; D002855:Chromatography, Thin Layer / Q000379:methods*; D005456:Fluorescent Dyes / Q000737:chemistry; D006147:Guanine / Q000031:analogs & derivatives / Q000737:chemistry; D006801:Humans; D009369:Neoplasms; D019853:O(6)-Methylguanine-DNA Methyltransferase / Q000032:analysis*
D005456:Fluorescent Dyes; C064976:O(6)-benzylguanine; D006147:Guanine; D019853:O(6)-Methylguanine-DNA Methyltransferase