Journal of East China Normal University(Natural Sc ›› 2012, Vol. 2012 ›› Issue (5): 45-53,84.

• Article • Previous Articles     Next Articles

Development of a luciferase-based method for rapid cell viability detection

ZHU Jing-jing, BAO Qiu-ying, WANG Yu-meng, XIA Gang   

  1. Key Laboratory of Brain Functional Genomics, Ministry of Education, Shanghai Key Laboratory of Brain Functional Genomics, East China Normal University, Shanghai 200062, China
  • Received:2011-06-01 Revised:2011-09-01 Online:2012-09-25 Published:2012-09-29

Abstract: The firefly luciferase gene was PCR-amplified from plasmid pGL4.26 and subcloned into a bacterial overexpression vector pET24a. Expressed luciferase fusion protein was purified using Ni-NTA affinity chromatograph and its activity was confirmed using Bright-GloTM kit. Based on the ATP-dependency of luciferase reaction, we developed a cell viability assay, which is faster, more convenient, and more sensitive in detect cell viability than generally used MTT, CCK-8 and Alamar Blue methods.

Key words: firefly luciferase, protein expression, protein purification, cell viability assay

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