Human Phosphofructokinase, Platelet ELISA Kit

From Antibodies.com, this sandwich ELISA is designed for the in vitro quantitative determination of human Phosphofructokinase, Platelet (PFKP) in tissue homogenates, cell lysates or other biological fluids. The assay uses a 96-well microtiter plate pre-coated with a PFKP-specific antibody. Standards and samples are added to the wells, where PFKP is captured by the immobilized antibody. After washing, a Biotinylated Anti-PFKP Antibody binds the captured PFKP, followed by HRP-Avidin conjugate. TMB substrate visualises the HRP enzymatic reaction, producing a blue product that turns yellow upon addition of acidic stop solution. Absorbance is read at 450 nm in a microplate reader and is proportional to PFKP concentration.

Key performance characteristics include a sensitivity of <0.136 ng/ml, an assay range of 0.312–20 ng/ml and a total assay time of 4h 30m. Spike-and-recovery validation across five replicates returned averages of 91% for serum (range 80%–102%), 90% for EDTA plasma (range 81%–100%) and 84% for heparin plasma (range 80%–89%). Dilution linearity across 1:2, 1:4, 1:8 and 1:16 dilutions was 87–91%, 87–107%, 74–101% and 92–97% respectively 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 include a pre-coated 96-well microplate (12 x 8 well strips), lyophilised standard, Detection Solution A, Detection Solution B, Wash Buffer (30X), Sample Dilution Buffer, TMB Substrate, Stop Solution and plate sealers. Components should be stored at +4°C or −20°C as instructed, and the kit should not be used past its expiration date.