A rapid β-NADH-linked fluorescence assay for lactate dehydrogenase in cellular death

Jeffery H. Moran, Rick G. Schnellmann

Research output: Contribution to journalArticlepeer-review

59 Scopus citations

Abstract

Lactate dehydrogenase (LDH) release is a common marker of cellular death. Traditionally, the fraction of LDH released has been measured using a NADH-linked W-Vis spectrophotometric method. The limitation of this method is that samples are usually run serially and thus is time intensive. Therefore, we developed a NADH-linked LDH assay using a fluorescence plate reader that had a correlation of 0.95 with the traditional UV-Vis spectrophotometric method. Using rabbit renal proximal tubule suspensions at a concentration of 1 mg cellular protein/ml of media, the fluorescence assay can determine LDH release in 22 samples in 2 min using 12 μL of cellular homogenates and 150 μL of media. The parallel processing of samples and smaller volumes used in the fluorescence assay results in decreased analysis time and costs.

Original languageEnglish (US)
Pages (from-to)41-44
Number of pages4
JournalJournal of Pharmacological and Toxicological Methods
Volume36
Issue number1
DOIs
StatePublished - Sep 1996
Externally publishedYes

Keywords

  • Apoptosis
  • Cell death
  • Fluorescence assay
  • Lactate dehydrogenase
  • Oncosis
  • Phrases
  • Renal cells

ASJC Scopus subject areas

  • Toxicology
  • Pharmacology

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