Abstract
LDH (Lactate dehydrogenase) enzyme catalyzes the reversible conversion of pyruvate to lactate using NAD+ as a cofactor. Although the physiological significance of lactate accumulation in tumor cells, a dead-end product in cellular metabolism, is currently a topic of debate, it has long been known that many tumor cells express a high level of LDH-A (Koukourakis et al., 2003; Koukourakis et al., 2006; Koukourakis et al., 2009). So detection of its enzyme activity in vitro is important for researching on LDH-A. Recently, it has been reported that Lys-5 acetylation could decrease LDH-A enzyme activity (Zhao et al., 2013).
Keywords: LDH, Enzyme activity, Lactate, NAD+
Materials and Reagents
Equipment
Procedure
Recipes
Acknowledgments
This protocol has been adapted from the previously published paper Zhao et al. (2013) and is described in further detail. We thank the members of the Fudan MCB laboratory for discussions throughout this study. We also thank Dr. Liming Wei for the IEF assay. This work was supported by the Chinese Ministry of Sciences and Technology 973 (Grant No. 2009CB918401, 2011CB910600), (Grant No. NCET-09-0315), NSFC (Grant No.31271454, 81225016) and NSFC-NIH (Grant No. 81110313). This work was also supported by Chinese Ministry of Education 985 Program, 100 Talents Program of Shanghai Health and Scholar of “Dawn” Program of Shanghai Education Commission and Shanghai Key basic research program(12JC1401100) to Q.Y.L. and NIH grants (Y.X. and K.L.G.); and the Fudan University Medical School Graduate Student Ming Dao Project Funds (D.Z.). This work is dedicated to the memory of Zhen Yu, who prepared the K5 acetylation antibody.
References
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