O(6)-alkylguanine is the major mutagenic and carcinogenic lesion in DNA induced by simple alkylating mutagens because of its preference for pairing with thymine during DNA replication. This adduct in DNA is removed by a ubiquitous and unique repair protein, O(6)-methylguanine-DNA methyltransferase (EC 2.1.1.63).Methylated-DNA--protein-cysteine methyltransferase, unlike true enzymes, accepts the alkyl group from the lesion in a stoichiometric second-order reaction. The methyl-acceptor residue is cysteine. Tano et al. (1990) cloned the cDNA of the human MGMT gene in an expression vector on the basis of its rescue of a methyltransferase-deficient E. coli host.
Human Methylated-DNA--protein-cysteine methyltransferase (MGMT) ELISA Kit employs a two-site sandwich ELISA to quantitate MGMT in samples. An antibody specific for MGMT has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and anyMGMT present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for MGMT is added to the wells. After washing, Streptavidin conjugated Horseradish Peroxidase (HRP) is added to the wells. Following a wash to remove any unbound avidin-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of MGMT bound in the initial step. The color development is stopped and the intensity of the color is measured.
Human Methylated-DNA--protein-cysteine methyltransferase (MGMT) ELISA Kit listed herein is for research use only and is not intended for use in human or clinical diagnosis. Suggested applications of our products are not recommendations to use our products in violation of any patent or as a license. We cannot be responsible for patent infringements or other violations that may occur with the use of this product.
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