Thin layer chromatography-based assay of O6-methylguanine-DNA methyltransferase activity in tissue

Cliff Robinson; Juan Palomo; Michael A Vogelbaum

doi:10.1016/j.ab.2010.06.019

Thin layer chromatography-based assay of O6-methylguanine-DNA methyltransferase activity in tissue

Anal Biochem. 2010 Oct 15;405(2):263-5. doi: 10.1016/j.ab.2010.06.019. Epub 2010 Jun 15.

Authors

Cliff Robinson¹, Juan Palomo, Michael A Vogelbaum

Affiliation

¹ Department of Radiation Oncology, Washington University in St. Louis, St. Louis, MO 63110, USA.

PMID: 20558126
DOI: 10.1016/j.ab.2010.06.019

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.

MeSH terms

Cell Line, Tumor
Chromatography, Thin Layer / methods*
Fluorescent Dyes / chemistry
Guanine / analogs & derivatives
Guanine / chemistry
Humans
Neoplasms
O(6)-Methylguanine-DNA Methyltransferase / analysis*

Substances

Fluorescent Dyes
O(6)-benzylguanine
Guanine
O(6)-Methylguanine-DNA Methyltransferase