Human Methylated-DNA--protein-cysteine methyltransferase (MGMT) ELISA Kit

SKU: AE58337HU

Size: 96T. Other sizes are also available.

Species Reactivity: Human (Homo sapiens)

UniProt: P29177

Abbreviation: MGMT

Alternative Names: RP11-109A6.1; O6-methylguanine-DNA methyltransferase|methylguanine-DNA methyltransferase

Application: ELISA

Range: 0.156-10 ng/mL

Sensitivity: 0.055 ng/mL

Test principle: This assay 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.

Product Overview: 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.

Stability: The stability of ELISA kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition. The loss rate was determined by accelerated thermal degradation test. Keep the kit at 37°C for 4 and 7 days, and compare O.D.values of the kit kept at 37°C with that of at recommended temperature. (referring from China Biological Products Standard, which was calculated by the Arrhenius equation. For ELISA kit, 4 days storage at 37°C can be considered as 6 months at 2 - 8°C, which means 7 days at 37°C equaling 12 months at 2 - 8°C).