Magnesium, manganese, and divalent cation transport assays in intact cells.
Protocols are described for the measurement of radioisotopic cation influx and efflux in bacteria using Salmonella as a model system. Methods are discussed for both the use of primary radioisotopes for measurement, e.g., using 54Mn2+ to measure Mn2+ influx, and the use of surrogate radioisotopes when the primary radioisotope is not available, e.g., use of 63Ni2+ or 57Co2+ in place of 28Mg2+ to measure Mg2+ influx. Both vacuum filtration and centrifugation assays are described. In addition, the use and misuse of chelating agents is discussed.
Maguire
Michael E
ME
Department of Pharmacology, Case School of Medicine, Case Western Reserve University, Cleveland, OH, USA.
D002021:Buffers; D002413:Cations, Divalent / Q000378:metabolism; D002614:Chelating Agents; D017136:Ion Transport / Q000235:genetics; D007700:Kinetics; D008024:Ligands; D008274:Magnesium / Q000378:metabolism*; D008345:Manganese / Q000378:metabolism*; D011868:Radioisotopes / Q000378:metabolism; D012475:Salmonella / Q000235:genetics / Q000378:metabolism*
D002021:Buffers; D002413:Cations, Divalent; D002614:Chelating Agents; D008024:Ligands; D011868:Radioisotopes; D008345:Manganese; D008274:Magnesium