Detection of necrosis by release of lactate dehydrogenase activity

Methods Mol Biol. 2013;979:65-70. doi: 10.1007/978-1-62703-290-2_7.

Abstract

Apoptosis and necrosis are two major forms of cell death observed in normal and disease pathologies. Although there are many assays for detection of apoptosis, relatively few assays are available for measuring necrosis. A key signature for necrotic cells is the permeabilization of the plasma membrane. This event can be quantified in tissue culture settings by measuring the release of the intracellular enzyme lactate dehydrogenase (LDH). When combined with other methods, measuring LDH release is a useful method for the detection of necrosis. In this chapter, we describe the step-by-step procedure for detection of LDH release from necrotic cells using a microtiter plate-based colorimetric absorbance assay.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biocatalysis
  • Cell Culture Techniques
  • Colorimetry / methods*
  • Electron Transport
  • Humans
  • L-Lactate Dehydrogenase / metabolism*
  • NAD / metabolism
  • Necrosis / enzymology*
  • Tetrazolium Salts / metabolism

Substances

  • Tetrazolium Salts
  • NAD
  • L-Lactate Dehydrogenase