Amplex UltraRed enhances the sensitivity of fluorimetric pyruvate detection.

ABSTRACT

Pyruvate, which is produced in the final step of glycolysis, participates in anabolic metabolism, catabolic metabolism, and signal transduction. We have recently reported a new sensitive and selective fluorimetric assay for pyruvate measurement using Amplex Red as a fluorogenic dye. However, the fluorescence of the reaction product, resorufin, is pH dependent, which limits the sensitivity of the assay at pH 6.7. In this Note, we evaluate Invitrogen's new dye, Amplex UltraRed, as a fluorogenic agent for pyruvate determination. Our results show that Amplex UltraRed improves the performance of the assay, providing brighter fluorescence and enhanced sensitivity.