Rat Protease Activated Receptor 4 ELISA Kit

From Antibodies.com, this sandwich enzyme-linked immunosorbent assay (sELISA) is designed for the in vitro quantitative determination of rat Protease Activated Receptor 4 (PAR4) in tissue homogenates, cell lysates and other biological fluids. The platform is a pre-coated 96-well microplate arranged as 12 x 8 well strips. Standards and test samples are added to the wells, where PAR4 is captured by the immobilised antibody. After washing, a Biotinylated Anti-PAR4 Antibody is added, followed by an HRP-Avidin conjugate. TMB substrate is then used to visualise the enzymatic reaction, producing a blue product that turns yellow upon addition of the acidic stop solution. The absorbance is read at 450 nm in a colorimetric microplate reader, and PAR4 concentration is calculated by reference to the standard curve.

The kit provides a quantitative assay range of 0.156–10 ng/ml with a sensitivity of <0.059 ng/ml, and the complete protocol takes 4 hours 30 minutes. Kit components are stored at +4 °C or −20 °C as instructed and should not be used past the expiration date. Included reagents comprise a lyophilised standard, detection solutions A and B, 30X wash buffer, sample dilution buffer, TMB substrate, stop solution and adhesive plate sealers.

Sample recovery was assessed in serum, EDTA plasma and heparin plasma (n=5 per matrix). For undiluted samples the average recovery was 91% in serum (range 80–102%), 90% in EDTA plasma (range 81–100%) and 84% in heparin plasma (range 80–89%). Across dilution factors of 1:2, 1:4, 1:8 and 1:16, recoveries fell within 74–107% for serum, 79–108% for EDTA plasma and 82–105% for heparin plasma.