Mouse DNA Repair Protein RAD50 ELISA Kit

This sandwich Enzyme-Linked Immunosorbent Assay (sELISA) from Antibodies.com is intended for the in vitro quantitative determination of mouse Rad50 in tissue homogenates, cell lysates or other biological fluids. The assay uses an antibody specific for Rad50 that has been pre-coated onto a 96-well microplate. Standards and test samples are added to the wells and any Rad50 present is captured by the immobilised antibody. After washing, a Biotinylated Anti-Rad50 Antibody is added, followed by an HRP-Avidin conjugate. A TMB substrate solution is then used to visualise the HRP enzymatic reaction, producing a blue-coloured product that turns yellow upon addition of acidic stop solution. The density of yellow is proportional to the amount of Rad50 captured, and the concentration is calculated by reading the O.D. absorbance at 450 nm in a microplate reader and referring to a standard curve.

The kit has a stated sensitivity of <0.113 ng/ml and an assay range of 0.312–20 ng/ml. It is read on a colorimetric microplate reader, with a total assay time of 4 h 30 min, and the components should be stored at +4 °C or -20 °C as instructed; the kit should not be used past its expiration date. Recovery has been evaluated in serum, EDTA plasma and heparin plasma samples (n=5), with average recoveries of 91%, 94% and 96% respectively. Dilution linearity was assessed at 1:2, 1:4, 1:8 and 1:16 in the same sample types. The kit, which targets the mouse DNA Repair Protein RAD50, is suitable for cell lysates and tissue homogenates as a quantitative sandwich assay.