Mouse Dihydroorotate Dehydrogenase ELISA Kit

Manufactured by Antibodies.com, this kit is designed for the in vitro quantitative determination of mouse Dihydroorotate Dehydrogenase (DHODH) in tissue homogenates or other biological fluids. It is configured as a sandwich ELISA for use with mouse samples and is run on a pre-coated 96-well microplate arranged as 12 x 8 well strips, with colourmetric detection read at 450 nm in a microplate reader.

The assay follows a sandwich enzyme immunoassay format. An antibody specific for DHODH is pre-coated onto the plate; standards and test samples are added and the target is captured by the immobilised antibody. After washing, a Biotinylated Anti-DHODH Antibody is added, followed by an HRP-Avidin conjugate. TMB substrate is then used to visualise the HRP-catalysed reaction, producing a blue product that turns yellow upon addition of an acidic stop solution. The intensity of the yellow colour is proportional to the amount of DHODH captured, and the concentration is calculated from the absorbance at 450 nm against the standard curve. The full assay time is 4h 30m.

Reported performance specifications include a sensitivity of <0.056 ng/ml and an assay range of 0.156–10 ng/ml. Spike-and-recovery data (n=5) are given for serum (80–102%, average 91%), EDTA plasma (81–100%, average 90%) and heparin plasma (80–89%, average 84%). Dilution linearity at 1:2, 1:4, 1:8 and 1:16 is reported as: serum 87–91%, 87–107%, 74–101% and 92–97%; EDTA plasma 90–105%, 84–101%, 90–101% and 79–108%; heparin plasma 84–95%, 92–105%, 82–105% and 89–91%. Kit components are stored at +4°C or -20°C as instructed, and the product should not be used past its expiration date.