Abstract | INTRODUCTION: Quantification of lactate dehydrogenase (LDH) release is a widely accepted assay for the quantitative determination of cell viability and late-stage apoptosis. Major disadvantages of commercially available LDH assay kits include proprietary formulations, limited options for optimization and high cost, all resulting in limited reproducibility in research applications. Here, we describe a novel, custom LDH assay suitable in the context of plate reader-based screening of drug candidates for glioprotection, but with wide applicability to other cell types and experimental paradigms. METHODS: RESULTS: There was no statistically significant difference between results obtained with the novel custom assay and a commercially available assay CytoTox96® ( Promega, Madison, WI). DISCUSSION: The novel custom LDH release assay allows the reproducible quantification of cell viability and is highly cost-effective when compared to commercially available assays (approximately 25 times cheaper). In addition and in contrast to commercially available assays, the identification and detailed description of all assay components and procedures provide greater control over experimental conditions and design. We provide a detailed standard operating procedure permitting our novel assay to be readily adapted depending on experimental requirements.
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Authors | Simon Kaja, Andrew J Payne, Tulsi Singh, Jasleen K Ghuman, Erin G Sieck, Peter Koulen |
Journal | Journal of pharmacological and toxicological methods
(J Pharmacol Toxicol Methods)
2015 May-Jun
Vol. 73
Pg. 1-6
ISSN: 1873-488X [Electronic] United States |
PMID | 25681780
(Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)
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Copyright | Copyright © 2015 Elsevier Inc. All rights reserved. |
Chemical References |
- Antioxidants
- Reactive Oxygen Species
- tert-Butylhydroperoxide
- L-Lactate Dehydrogenase
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Topics |
- Antioxidants
(pharmacology)
- Cell Line, Tumor
- Cell Survival
(drug effects, physiology)
- Drug Evaluation, Preclinical
(methods)
- Humans
- L-Lactate Dehydrogenase
(antagonists & inhibitors, metabolism)
- Neurosciences
- Reactive Oxygen Species
(metabolism)
- tert-Butylhydroperoxide
(pharmacology)
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