a-Ketoglutarate Detection Assay Kit

Cayman’s a-KG Detection Assay provides fluorometric- or colorimetric-based methods for quantifying a-KG in biological samples such as serum, plasma, urine, and tissue. It can also be utilized to determine intracellular and extracellular a-KG concentrations in cell culture samples. In this assay, alanine transaminase (ALT) deaminates a-KG to form pyruvate.{33073,33074} Pyruvate is then oxidized by pyruvate oxidase to yield acetyl phosphate, hydrogen peroxide (H2O2), and carbon dioxide.{33075} As H2O2 is produced, the cycling mechanism of horseradish peroxidase (HRP) results in the concurrent reduction of H2O2 to H2O and the oxidation of 10-acetyl-3,7-dihydroxyphenoxazine (ADHP) to produce the highly fluorescent compound resorufin.{33076} Resorufin fluorescence is analyzed with an excitation wavelength between 530-540 nm and an emission wavelength between 585-595 nm. Alternatively, the absorbance of resorufin can be measured at 570 nm.