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Analysis of Cell Viability by the Lactate Dehydrogenase Assay
Abstract
A common method for determining cytotoxicity is based on measuring the activity of cytoplasmic enzymes released by damaged cells. Lactate dehydrogenase (LDH) is a stable cytoplasmic enzyme that is found in all cells. LDH is rapidly released into the cell culture supernatant when the plasma membrane is damaged, a key feature of cells undergoing apoptosis, necrosis, and other forms of cellular damage. LDH activity can be easily quantified by using the NADH produced during the conversion of lactate to pyruvate to reduce a second compound in a coupled reaction into a product with properties that are easily quantitated. This protocol measures the reduction of a yellow tetrazolium salt, INT, by NADH into a red, water-soluble formazan-class dye by absorbance at 492 nm. The amount of formazan is directly proportional to the amount of LDH in the culture, which is, in turn, directly proportional to the number of dead or damaged cells.
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Cited by
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Zhong, D., Wang, R., Zhang, H., Wang, M., Zhang, X., & Chen, H. (2023) · Nature Communications · doi:10.1038/s41467-023-39716-7 · Cited by 52
In vitro Liver Zonation of Primary Rat Hepatocytes
Tomlinson, L., Hyndman, L., Firman, J. W., Bentley, R., Kyffin, J. A., Webb, S. D., McGinty, S., & Sharma, P. (2019) · Frontiers in Bioengineering and Biotechnology · doi:10.3389/fbioe.2019.00017 · Cited by 38
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Metadata
- Authors
- Kumar, P., Nagarajan, A., & Uchil, P. D.
- Venue
- Cold Spring Harbor Protocols
- Publisher
- Cold Spring Harbor Laboratory
- Published
- 2018-06
- Volume
- 2018
- Issue
- 6
- Pages
- pdb.prot095497
- ISSN
- 1559-6095 (ISSN-L)
- 1940-3402
- OpenAlex
- W2807227439
- Cited by
- 666
- References
- 0
- Type
- posted content
- Language
- en
- Open access
- closed
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