L-Lactate Assay- Fluorescence

Lactate is an important intermediary in glucose metabolism. Under hypoxic or anaerobic conditions, lactate dehydrogenase (LDH) converts pyruvate, the final product of glycolysis, to lactate. Monitoring lactate levels is an adequate indicator of the balance between tissue oxygen demand and utilization, and is useful when studying cellular physiology. The L-lactate assay relies on the LDH-catalyzed oxidation of NAD to NADH. In a coupled reaction, the generated NADH reduces a probe, producing a highly fluorescent product. The fluorescence intensity, measured at an excitation wavelength of 560 nm and an emission wavelength of 590 nm, is directly proportional to the lactate concentration in the sample. The assay demonstrates a linear response up to 50 µM L-lactate, with a detection limit as low as 1 µM.