Microplate Assays for Spectrophotometric Measurement of Mitochondrial Enzyme Activity

Methods Mol Biol. 2019:1978:355-368. doi: 10.1007/978-1-4939-9236-2_22.

Abstract

Spectrophotometric analysis of metabolic enzyme activity from homogenized tissues is a valuable method for investigating mitochondrial content and capacity. Enzyme activity is normally measured in single cuvette spectrophotometers, requiring a large sample volume and low throughput. Here, we describe microplate assays for high-throughput analysis of mitochondrial enzymes citrate synthase, β-hydroxyacyl CoA dehydrogenase, aconitase, and mitochondrial electron transport system (ETS) complexes I, II, III, and IV.

Keywords: Citrate synthase; Electron transport chain; High throughput; Mitochondrial complexes; Mitochondrial function.

MeSH terms

  • Aconitate Hydratase / chemistry
  • Electron Transport / genetics
  • Electron Transport Complex I / chemistry*
  • Enzymes / chemistry*
  • Enzymes / metabolism
  • Mitochondria / chemistry
  • Mitochondria / enzymology*
  • Spectrophotometry / methods*

Substances

  • Enzymes
  • Aconitate Hydratase
  • Electron Transport Complex I