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Interference of electron donors in the measurement of the proton gradient and membrane potential by flow dialysis
Authors:O H Setty  R W Hendler
Affiliation:Laboratory of Cell Biology, National Heart, Lung, and Blood Institute, National Institutes of Health, Building 3, Room B1-22, Bethesda, MD 20205, U.S.A.
Abstract:
The three most commonly used electron donors for flow dialysis measurements of membrane potential lead to the development of an apparent but artifactual membrane potential with the interior negative in the presence or absence of membrane vesicles. The same three electron donors used in flow dialysis determinations of delta pH in the presence or absence of membrane vesicles lead to the development of an apparent but artifactual delta pH with the interior acidic. These artifacts have been evaluated using two probes for membrane potential, namely, TPP+ and rubidium in the presence of valinomycin and for two probes of delta pH, namely, acetate and DMO. Measurements were made over a range of ionic strengths.
Keywords:flow dialysis  proton gradient  membrane potential  electron donor  FCCP  DMO  5,5′-dimethyl-oxazolidine-2,4-dione  tetraphenylphosphonium (bromide salt)  PMS  phenazine methosulfate  ΔpH  transmembrane pH difference  Δψ  transmembrane difference in electrical potential
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