TCA Cycle Assays
Assay every step of the TCA (Krebs, citric acid) cycle: the intermediates from citrate to oxaloacetate, the enzymes that convert them, and the ATP and NADH the cycle produces.
Citrate, isocitrate, alpha-ketoglutarate, succinate, fumarate, malate and oxaloacetate, plus pyruvate and acetyl-CoA.
Citrate synthase, aconitase, isocitrate dehydrogenase, alpha-KGDH, succinate dehydrogenase, fumarase, malate dehydrogenase and PDH.
ATP, ADP, the ADP/ATP ratio and NAD/NADH.
The TCA cycle, also known as the Krebs or citric acid cycle, is the central pathway of aerobic energy metabolism. It runs in the mitochondria as eight enzyme-catalysed reactions that oxidise acetyl-CoA to carbon dioxide while generating NADH, FADH2 and ATP or GTP.
The cycle also supplies building blocks for amino acid, lipid and nucleotide synthesis, so changes in its intermediates are informative in cancer metabolism, immunometabolism and mitochondrial disease. Inherited defects in TCA cycle enzymes are linked to conditions such as Leigh syndrome and paraganglioma.
Yes. They are three names for the same pathway, first described by Hans Krebs in 1937.
Metabolite assays show where intermediates accumulate or run low. Enzyme activity assays show which step is impaired. Using both locates a defect more precisely.
Depending on the kit: cells, tissue homogenates, isolated mitochondria, serum and plasma.