Phosphoenolpyruvate Assay Kit

Cell Biolabs’ Phosphoenolpyruvate Assay Kit measures total PEP within biological samples.  PEP in the presence of ADP is converted by Pyruvate Kinase to pyruvate + adenosine triphosphate (ATP). Pyruvate is converted by pyruvate oxidase in the presence of phosphate and oxygen into acetyl phosphate, carbon dioxide, and hydrogen peroxide.  The resulting hydrogen peroxide is then detected with a highly specific fluorometric probe.  Horseradish peroxidase catalyzes the reaction between the probe and hydrogen peroxide, which bind in a 1:1 ratio.  Samples are compared to a known concentration of PEP standard within the 96-well microtiter plate format.  Samples and standards are incubated for 30 minutes and then read with a standard 96-well fluorometric plate reader.