Van Bergen 2014 Mitochondrion
|Van Bergen NJ, Blake RE, Crowston JG, Trounce IA (2014) Oxidative phosphorylation measurement in cell lines and tissues. Mitochondrion 15:24-33.|
Abstract: Mitochondrial oxidative phosphorylation (OXPHOS) dysfunction is implicated in a growing spectrum of diseases, from neurodegeneration to cancer. Where tissues or transformed cells are available, respirometry and enzymology allow a sophisticated analysis of OXPHOS with modest-cost equipment. The isolation of organelle fractions is also invaluable for determining association of proteins of interest. Here we revisit and consolidate methods to measure whole cell mitochondrial ATP synthesis, respiration, isolation of mitochondria from cultured cells and tissues, and OXPHOS enzymology. We also explain common pitfalls, guide optimisation of the methods for new users, and provide full laboratory protocols in Supplementary materials.
• O2k-Network Lab: AU Melbourne Trounce IA
Labels: MiParea: Respiration, Instruments;methods, mtDNA;mt-genetics, mt-Medicine
Organism: Human, Mouse Tissue;cell: Nervous system, Blood cells, Lymphocyte Preparation: Intact cells, Permeabilized cells, Isolated mitochondria Enzyme: Complex I, Complex II;succinate dehydrogenase, Complex III, Complex IV;cytochrome c oxidase, Marker enzyme Regulation: ATP production, Uncoupler Coupling state: LEAK, ROUTINE, OXPHOS, ET Pathway: N, S, NS HRR: Oxygraph-2k, TIP2k, O2k-Protocol
Blood cell preparation