Rat O-6-Methylguanine DNA Methyltransferase ELISA Kit

From Antibodies.com, this sandwich ELISA (sELISA) is designed for the in vitro quantitative determination of O-6-Methylguanine DNA Methyltransferase (MGMT) in rat tissue homogenates or other biological fluids. An antibody specific for MGMT is pre-coated onto a 96-well microtiter plate. Standards and test samples are added to the wells and MGMT present in each sample is bound by the immobilized antibody. Following incubation and washing, a Biotinylated Anti-MGMT Antibody is added, which binds the captured MGMT. An HRP-Avidin conjugate is then added and, after a further incubation and washing step, TMB substrate solution is used to visualise the enzymatic reaction, producing a blue product that turns yellow upon addition of acidic stop solution. The density of yellow is proportional to the amount of MGMT captured, and the concentration is calculated by reading the O.D. absorbance at 450 nm in a microplate reader and referring to the standard curve.

The assay delivers a sensitivity of <27 pg/ml and a quantitative range of 78.1-5000 pg/ml, with a total assay time of 4h 30m and colorimetric detection on a pre-coated microplate platform. Reported recovery values (n=5) are: serum 80%-102% (average 91%), EDTA plasma 81%-100% (average 90%), and heparin plasma 80%-89% (average 84%). Kit components should be stored at +4°C or -20°C as instructed, and the product should not be used past its expiration date.