Dehydrogenase-Glo™ Detection System

Measuring dehydrogenase activity in K562 cells. K562 cells were diluted twofold starting at 2.5 × 10⁴ cells/ml in PBS, and 50μl was transferred into a 96-well assay plate. Dehydrogenase detection reagent was prepared for each dehydrogenase, and 50μl was added to sample wells. No-substrate controls (NAD only and NADP only) were included to determine background from nonspecific enzyme activity in K562 cells. Panel A. Luminescence was measured for dehydrogenases and controls after 60 minutes at different cell densities. Panel B. Time-dependent increase in GAPDH activity was shown at different cell densities. Luminescence was read every 15 minutes for 120 minutes. Solid black lines indicate linear range for each cell number. Panel C. Specific activity for each dehydrogenase was calculated. Data represent the average of three replicates with error bars indicating one standard deviation.