Automated fluorometric analysis of DNA, protein, and enzyme activities: application to methods in cell culture

Automated fluorometric methods for the analysis of DNA, protein, and selected enzyme activities for N-acetyl-β-d-glucosaminidase, glucose 6-phosphate dehydrogenase, and 6-phosphogluconate dehydrogenase are described. Instrumentation for these assays includes a Gilford 3500 computer-directed analyzer in conjunction with a Farrand Ratio Fluorometer-2 modified for flowthrough sampling. Comparisons were made between the automated fluorometric methods described and manual spectrophotometric or fluorometric methods for reproducibility, speed of analysis, and quantitative correlation. Typical values of N-acetyl-β-d-glucosaminidase, glucose 6-phosphate dehydrogenase, and 6-phosphogluconate dehydrogenase activities obtained by these methods in isolated rat hepatocytes and Reuber H-35 hepatoma cells are reported.