Measurement of oxygen consumption was performed with a Clark-type oxygen electrode system (Hansatech Instruments, Norfolk, UK).54 (link) In total, 4 × 106 cells were suspended in air-saturated buffer E (300 mM mannitol; 5 mM MgCl2; 10 mM KCl; 10 mM KH2PO4, pH 7.4; and 1 mg/ml bovine serum albumin, fatty acid free) and placed into a water-jacketed chamber at 37 °C. Oxygen consumption under conditions of chemical inhibition of the respective RC complexes was recorded, related to total cell number and expressed as femtomoles O2 per cell per min.
Clark type oxygen electrode system
Manufactured by Hansatech
Sourced in United Kingdom
The Clark-type oxygen electrode system is a laboratory instrument used to measure the concentration of dissolved oxygen in a sample. It functions by utilizing an electrochemical cell that generates a current proportional to the amount of oxygen present in the sample. The system provides a quantitative measurement of oxygen levels, enabling researchers to monitor and analyze oxygen-dependent processes.
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Lab products found in correlation
2 protocols using clark type oxygen electrode system
1
Measurement of Cellular Oxygen Consumption
2
Mitochondrial Respiration Profiling
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Mitochondrial
respiration rates (state 3, ADP-stimulated; state 4, ADP-deleted;
and respiratory control ratio) were observed by a Clark-type oxygen
electrode system connected to a computer-operated oxygraph control
unit (Hansatech Instruments, Norfolk, U.K.).41 (link) About 1.0 mL of total reaction volume was used at 28 °C for
all of the experiment group conditions. About 30–60 μg
of freshly isolated mitochondria was added to the respiration buffer
(120 mM KCl, 5 mM potassium phosphate, 3 mM HEPES, 1 mM ethylene glycol-O,-O′-bis-(2-aminoethyl) tetraacetic
acid (EGTA), 1 mM MgCl2, and 0.2% BSA, pH 7.2) and allowed
to equilibrate for a minute. FAD-linked substrate (10 mM succinate)
was then added to the chamber and allowed to equilibrate for 1 min,
followed by the addition of ADP (100 μM final concentration).
The measurements involve addition of ADP for state 3 and removal of
ADP for state 4. The respiratory control ratio (RCR) was calculated
by the ratio of state 3 to state 4 respirations.42 (link)
respiration rates (state 3, ADP-stimulated; state 4, ADP-deleted;
and respiratory control ratio) were observed by a Clark-type oxygen
electrode system connected to a computer-operated oxygraph control
unit (Hansatech Instruments, Norfolk, U.K.).41 (link) About 1.0 mL of total reaction volume was used at 28 °C for
all of the experiment group conditions. About 30–60 μg
of freshly isolated mitochondria was added to the respiration buffer
(120 mM KCl, 5 mM potassium phosphate, 3 mM HEPES, 1 mM ethylene glycol-O,-O′-bis-(2-aminoethyl) tetraacetic
acid (EGTA), 1 mM MgCl2, and 0.2% BSA, pH 7.2) and allowed
to equilibrate for a minute. FAD-linked substrate (10 mM succinate)
was then added to the chamber and allowed to equilibrate for 1 min,
followed by the addition of ADP (100 μM final concentration).
The measurements involve addition of ADP for state 3 and removal of
ADP for state 4. The respiratory control ratio (RCR) was calculated
by the ratio of state 3 to state 4 respirations.42 (link)
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