Use of cellular glucose-6-phosphate dehydrogenase for cell quantitation: applications in cytotoxicity and apoptosis assays.
A fluorescence-based microplate assay was developed to quantify cell death based upon the measurement of glucose-6-phosphate dehydrogenase (G6PD) activity. G6PD is a cytosolic enzyme and leaks from cells when plasma membrane integrity is compromised. In this assay, cell death is measured by correlating the activity of extracellular G6PD to the reduction of resazurin to the fluorescent product, resorufin, via a coupled-enzyme reaction. The coupled-enzyme reaction permits rapid signal amplification from small amounts of G6PD, an advantage over assays based on resazurin alone. This assay is rapid, nontoxic, and amenable to high-throughput screening. The assay has a Z' factor of 0.78.
Batchelor
Robert H
RH
Molecular Probes, Inc., 29851 Willow Creek Road, Eugene, OR 97402, USA. rob.batchelor@probes.com
Zhou
Mingjie
M
D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D002166:Camptothecin / Q000633:toxicity; D002452:Cell Count / Q000295:instrumentation / Q000379:methods*; D002460:Cell Line; D005453:Fluorescence; D005954:Glucosephosphate Dehydrogenase / Q000097:blood / Q000378:metabolism*; D006801:Humans; D019169:Jurkat Cells; D051379:Mice; D010078:Oxazines / Q000032:analysis; D013379:Substrate Specificity; D014409:Tumor Necrosis Factor-alpha / Q000633:toxicity; D014466:Ultraviolet Rays / Q000009:adverse effects; D014966:Xanthenes / Q000032:analysis
D010078:Oxazines; D014409:Tumor Necrosis Factor-alpha; D014966:Xanthenes; C005843:resazurin; D005954:Glucosephosphate Dehydrogenase; D002166:Camptothecin