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

The Assay Genie bovine MGMT sandwich ELISA provides quantitative colorimetric detection of methylated-DNA--protein-cysteine methyltransferase in bovine serum and plasma. The 96-well microplate format uses pre-coated capture antibodies and matched detection antibodies to generate a standard curve, with absorbance typically read at 450 nm on a standard plate reader. Researchers in veterinary oncology and DNA repair biology apply this kit to evaluate MGMT protein concentrations as an indicator of alkylation damage response and chemoresistance mechanisms in cattle.

Each kit supplies reagents for one 96-well plate, supporting up to 96 assays when run in duplicate. Serum and plasma samples should be processed promptly or stored refrigerated at 2-8 degrees C; reconstituted standards and antibodies are commonly aliquoted and frozen at -20 degrees C to limit freeze-thaw cycles and protect from light. The protocol follows a standard sandwich ELISA workflow with incubation steps, wash cycles, and a TMB substrate reaction terminated with stop solution. For research use only.

MGMT (O6-methylguanine-DNA methyltransferase) is a direct, single-step DNA repair enzyme that transfers alkyl groups from the O6 position of guanine onto a conserved active-site cysteine residue, restoring base integrity without requiring any cofactor. In bovine cancer biology, MGMT promoter methylation and protein abundance are routinely measured as biomarkers of alkylator responsiveness and tumor suppressor status, with particular relevance to bovine lymphosarcoma, enzootic bovine leukosis, and hepatocellular carcinoma studies.

This Assay Genie kit is supplied as a pre-coated 96-well microplate paired with lyophilized recombinant standards, a biotinylated detection antibody, streptavidin-HRP conjugate, TMB substrate, stop solution, and a wash buffer concentrate. The calibration curve typically spans 0.156-10 ng/mL with intra-assay CV below 10% and inter-assay CV under 12%. Beyond serum and plasma, cell culture supernatants, tissue homogenates prepared from liver or lymph node biopsies, and skimmed milk whey have been reported as additional compatible matrices following in-house validation by the end user. Reconstituted standards and antibody vials should be aliquoted and stored at -20 degrees C to limit freeze-thaw cycles and protected from light until first use.