Rat Phosphofructokinase, Muscle ELISA Kit

From Antibodies.com, this sandwich Enzyme-Linked Immunosorbent Assay is designed for the in vitro quantitative determination of rat Phosphofructokinase, Muscle (PFKM) in tissue homogenates and other biological fluids. The assay uses a 96-well microplate pre-coated with an antibody specific for PFKM. Standards and samples are added to the wells and PFKM is captured by the immobilised antibody. After washing, a biotinylated anti-PFKM antibody binds the captured target, followed by an HRP-avidin conjugate. A TMB substrate is added to visualise the enzymatic reaction, producing a blue product that turns yellow upon addition of acidic stop solution; absorbance is read at 450 nm and is proportional to PFKM concentration.

The working assay range is 0.312–20 ng/ml with a sensitivity of 0.127 ng/ml. Total assay time is 4 hours 30 minutes and detection is performed on a colorimetric microplate reader. Validation data report mean recoveries of 91% for serum, 90% for EDTA plasma, and 84% for heparin plasma (n=5), with linearity demonstrated across dilutions of 1:2 to 1:16. Kit components are stored at +4°C or −20°C as indicated.