Porcine C Reactive Protein ELISA Kit

From Antibodies.com, this sandwich Enzyme-Linked Immunosorbent Assay (ELISA) kit is intended for the in vitro quantitative determination of porcine C Reactive Protein in serum, plasma, tissue homogenates, cell lysates, cell culture supernates, and other biological fluids.

The assay uses a pre-coated 96-well microplate and employs the sandwich enzyme immunoassay technique. An antibody specific for C Reactive Protein is immobilised on the plate; standards and test samples are added and the target protein is captured by the immobilised antibody. After washing, a Biotinylated Anti-C Reactive Protein 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 acidic stop solution. The density of yellow is proportional to the amount of C Reactive Protein captured, and the concentration is calculated from the O.D. absorbance read at 450 nm on a colorimetric microplate reader.

The manufacturer reports a quantitative detection range of 0.156–10 ng/ml and a sensitivity of <0.064 ng/ml, with a total assay time of 4 h 30 min. Spike-and-recovery performance across matrices (n=5 each) was 80–102% (average 91%) for serum, 81–99% (average 90%) for EDTA plasma, and 80–89% (average 84%) for heparin plasma. Dilution linearity at 1:2, 1:4, 1:8 and 1:16 was reported as 87–91%, 87–107%, 74–101% and 92–97% for serum (n=5); 90–105%, 84–101%, 90–101% and 79–108% for EDTA plasma (n=5); and 84–95%, 92–105%, 82–105% and 89–91% for heparin plasma (n=5). Kit components should be stored at +4°C or -20°C as instructed and the kit should not be used past its expiration date.