Rat Pyruvate Kinase, Muscle ELISA Kit

A sandwich Enzyme-Linked Immunosorbent Assay (sELISA) from Antibodies.com, intended for the in vitro quantitative determination of rat Pyruvate Kinase Isozyme M1 in serum, plasma, tissue homogenates or other biological fluids. The assay uses a pre-coated microtiter plate with an antibody specific for Pyruvate Kinase Isozyme M1. Standards and samples are added to the wells, the target is captured by the immobilised antibody, and detection is achieved with a Biotinylated Anti-Pyruvate Kinase Isozyme M1 Antibody followed by an HRP-Avidin conjugate. TMB substrate is used to visualise the enzymatic reaction, producing a blue product that turns yellow on addition of acidic stop solution; absorbance is read at 450 nm in a microplate reader and the concentration is calculated from the standard curve.

The assay has a reported working range of 0.781–50 ng/ml and a sensitivity of 0.35 ng/ml, with a total assay time of 4h 30m. It is intended for use with a colorimetric microplate reader. Reported spike-recovery values were 80–102% (average 91%) in serum (n=5), 81–100% (average 90%) in EDTA plasma (n=5) and 80–89% (average 84%) in heparin plasma (n=5). Linearity across 1:2, 1:4, 1:8 and 1:16 dilutions was 87–91% / 87–107% / 74–101% / 92–97% for serum, 90–105% / 84–101% / 90–101% / 79–108% for EDTA plasma and 84–95% / 92–105% / 82–105% / 89–91% for heparin plasma (n=5). Kit components should be stored at +4°C or –20°C as instructed and the kit must not be used past its expiration date.