MTH1 Overexpression Lysate

HEK293T cells in 10-cm dishes were transiently transfected with a non-lipid polymer transfection reagent specially designed and manufactured for large volume DNA transfection. Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix, 1mM PMSF and 1mM Na3VO4, and then centrifuged to clarify the lysate. Protein concentration was measured by BCA protein assay kit.

Oxygen radicals damage chromosomal DNA causing cell death and inducing mutations. Among the various classes of DNA damage caused by oxygen radicals, an oxidized form of guanine base (8-oxoguanine) appears to be important as it can pair with cytosine and adenine and G:C to T:A transversion mutation occurs. A significant amount of 8-OxoG is formed in the chromosomal DNA of mammalian cells, with most damaged nucleotides excised from the DNA and excreted in the urine. Along with 8-oxoG being present in oxidatively damaged DNA, 8-oxo-deoxyguanosine (8-oxo-dGTP) is formed in the nucleotide pool during normal cellular metabolism and following oxidative stress. The 8-oxo-dGTP nucleotide can be incorporated in DNA during polymerization and can result in a mispairing unless repaired. MTH converts 8-oxo-dGTP in the nucleotide pool to the monophosphate and prevents the misincorporation of 8-oxo-dGTP into DNA (Figure courtesy of Dr. Mark Kelley). MTH also recognizes 8-oxo-rGTP, which could incorporate into RNA during gene transcription leading to missense or nonsense protein production.

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are guaranteed for 6 months from date of receipt.

Array NBP2-04636

• MTH1 Antibodies
• MTH1 Lysates
• MTH1 Proteins