Pyruvate Assay Kit

• MSDS: View or download

Cayman’s Pyruvate Assay provides both a fluorescence- and absorbance-based methods for quantifying pyruvate in biological samples such as serum, plasma, blood, urine, and saliva. It can also be utilized to determine intracellular and extracellular pyruvate concentrations in cell culture samples. In the assay, pyruvate oxidase catalyzes the conversion of pyruvate to acetyl phosphate, hydrogen peroxide (H2O2), and carbon dioxide. In the presence of horseradish peroxidase (HRP), H2O2 reacts stoichiometrically with 10-acetyl-3,7,-dihydroxyphenoxazine (ADHP) to produce the compound resorufin. Resorufin fluorescence is analyzed with an excitation wavelength between 530-540 nm and an emission wavelength between 585-595 nm. Resorufin absorbance is measured at a wavelength of 570 nm. This assay offers the option to measure absorbance or fluorescence. It is at the user’s discretion to choose the mode of measurement (and corresponding standard curve preparation) that best fits their needs. When read fluorometrically, this assay has a range of 0-75 µM and a lower limit of quantification (LLOQ) of 1.5 µM. When read colorimetrically, the range is 0-150 µM with an LLOQ of 3.0 µM.