Human Proteinase-activated receptor 4/PAR4 ELISA Kit

From Assay Genie, this sandwich ELISA, double antibody kit enables quantitative measurement of proteinase-activated receptor 4 (PAR4) in human serum, plasma, cell culture supernatant, cell or tissue lysate and other liquid samples. PAR4 is recognised as a regulator of platelet activation and function, making this assay relevant to cardiovascular, thrombosis and coagulation research. Synonyms: F2RL3, Coagulation factor II receptor-like 3, Thrombin receptor-like 3 (UniProt Q96RI0).

Detection is colorimetric, with a sensitivity of 0.469 ng/mL and a working range of 0.781-50 ng/mL. Linearity at 1:2, 1:4 and 1:8 dilutions (n = 5) was 84-101%, 86-101% and 85-101% in serum; 84-99%, 83-90% and 80-92% in EDTA plasma; and 89-103%, 87-103% and 85-104% in heparin plasma. Recovery ranges were 89-104% (serum, average 97%), 89-102% (EDTA plasma, average 97%) and 91-104% (heparin plasma, average 98%).

The protocol covers reagent and plate preparation, primary incubation at 37°C for 90 minutes, biotinylated detection antibody incubation at 37°C for 60 minutes, HRP-Streptavidin (SABC) binding at 37°C for 30 minutes, TMB colour development in the dark for 10–20 minutes, and absorbance reading at 450 nm after stop solution. Sample preparation guidance is provided for serum (clot, centrifuge 1000 × g for 20 minutes), plasma (anticoagulant tubes, 1000 × g for 15 minutes at 2–8°C), tissue homogenate, cell culture supernatant (2500 rpm for 5 minutes) and cell lysate. Supplied components include a dismountable ELISA microplate, lyophilised standard, 100X biotin-labelled antibody, 100X HRP-Streptavidin conjugate, TMB substrate, sample/antibody/SABC dilution buffers, 25X wash buffer, stop solution, plate sealer and technical manual. Storage: 2-8°C for 12 months.