title: Thin layer chromatography-based assay of O6-methylguanine-DNA methyltransferase activity in tissue.
abstract: 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.
authors.affiliation: Department of Radiation Oncology, Washington University in St. Louis, St. Louis, MO 63110, USA.
authors.forename: Cliff
authors.initials: C
authors.lastname: Robinson
authors.forename: Juan
authors.initials: J
authors.lastname: Palomo
authors.forename: Michael A
authors.initials: MA
authors.lastname: Vogelbaum
mesh_terms: 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*
chemical_list: D005456:Fluorescent Dyes; C064976:O(6)-benzylguanine; D006147:Guanine; D019853:O(6)-Methylguanine-DNA Methyltransferase