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O2 + 2 H2O + 2 e-+ → H2O2 + 2 OH- Electron flow to oxygen as a result of oxidative phosphorylation can be demonstrated using an oxygen electrode, ie. measuring the oxygen consumption activity of freshly isolated mitochondria. The electrode compartment is isolated from the reaction chamber (where the mitochondria and pulses, fuels etc. are added) by a thin Teflon membrane; the membrane is permeable to molecular oxygen and allows this gas to reach the cathode, where it is electrolytically reduced. The reduction allows a current to flow; this is converted to a potential and recorded on a flatbed chart recorder. The trace is thus a measure of the oxygen activity of the reaction mixture, and both the extent and rate of oxygen uptake by mitochondria may be measured. The current flowing is proportional to the activity of oxygen provided the solution is stirred constantly (machanical flea) to minimize the formation of an unstirred layer next to the membrane. Reference: Trinity College Dublin, Biochemistry Laboratory Manual for Senior Freshman Science, 2005-2006. www.tcd.ie/biochemistry |
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