Human Ubiquitin Conjugating Enzyme E2A ELISA Kit

A sandwich enzyme-linked immunosorbent assay (sELISA) from Antibodies.com, intended for the in vitro quantitative determination of human Ubiquitin Conjugating Enzyme E2A (also referred to as Rad6) in serum, plasma, tissue homogenates, cell lysates, cell culture supernates or other biological fluids. Reactivity is specific to human samples.

An antibody specific for Rad6 is pre-coated onto a 96-well microtiter plate. Standards and test samples are added and the target present in each sample is bound by the immobilised antibody. Following incubation and washing, a Biotinylated Anti-Rad6 Antibody is added, followed by an HRP-Avidin conjugate. TMB substrate is then used to visualise the HRP enzymatic reaction, producing a blue-coloured product that changes to yellow upon addition of acidic stop solution. Absorbance is read at 450 nm on a colorimetric microplate reader and the concentration of Rad6 is calculated by reference to the standard curve.

The assay reports a sensitivity of 0.081 ng/ml and a quantitative range of 0.312–20 ng/ml, with a total assay time of approximately 4 h 30 min. Recovery has been demonstrated in serum (average 91%), EDTA plasma (average 96%) and heparin plasma (average 94%), and dilution linearity has been characterised across sample dilutions of 1:2 to 1:16 in each of these matrices.