MGMT Assay Kit 

The kit can be used for:
• Screening for MGMT inhibitor.
• Detection of MGMT activity in cell lines.

MGMT (O⁶-Methylguanine-DNA methyltransferase) is a ubiquitous DNA repair protein that removes O⁶-alkyl-guanine lesions, primarily O⁶-methyl-guanine from damaged DNA. It is a major contributor to cellular protection from the mutagenic, carcinogenic, and cytotoxic effects of DNA alkylation.
The MGMT assay kit is based on the formation of a new restriction site in a 23-mer ³²P labeled ds-oligonucleotide after removal of the O⁶-methyl-guanine lesion. Cleavage of the new restriction site results in two smaller oligonucleotides that can be resolved on a denaturing PAGE gel.

sufficient for 100 tests

wet ice

−20°C

Hansen, W.K., and Kelly, M.R., Review of mammalian DNA repair and translational implications. J. Pharmacol. Exp. Ther. 295, 1-9, (2000) abstract

Margison, G.P., and Santibanez-Koref, M.F., O⁶-alkylguanine-DNA alkyltransferase: Role in carcinogenisis and chemotherapy. Bioessays 24, 255-266, (2002) abstract

Sassanfar, M., et al., Relative efficiencies of the bacterial, yeast, and human DNA methlytransferases for the repair of O⁶-methylguanine and O⁴-methylthymine. Suggestive evidence for O⁴-methylthymine repair by eukaryotic methyltransferases. J. Biol. Chem. 266, 2767-2771, (1991) abstract

Encell, L.P., and Loeb, L.A., Redesigning the substrate specificity of human O⁶-alkylguanine-DNA transferase. Mutants with enhanced repair of O⁴-methylthymine. Biochemistry 38, 12097-12103, (1999) abstract

UN 3316 9/PG 3

MGMT substrate O⁶-methylguanine strand 1 vial

MGMT substrate complementary strand 1 vial

Reaction buffer 10X 1.5 mL

Stop solution 1 mL

Methylguanine DNA methyltransferase (MGMT) 10 μg

O⁶Benzylguanine solution 50 μL

Spin column-10 2 units