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

Cliff Robinson, Juan Palomo, Michael A. Vogelbaum

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8 Scopus citations

Abstract

Expression of the DNA repair enzyme O⁶-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 O⁶-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.

Original language	English
Pages (from-to)	263-265
Number of pages	3
Journal	Analytical Biochemistry
Volume	405
Issue number	2
DOIs	https://doi.org/10.1016/j.ab.2010.06.019
State	Published - Oct 2010

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abstract = "Expression of the DNA repair enzyme O6-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 O6-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.",

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AB - Expression of the DNA repair enzyme O6-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 O6-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.

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

Abstract

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