1976
DOI: 10.1038/259047a0
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Use of ionophore A23187 to measure cytoplasmic Ca buffering and activation of the Ca pump by internal Ca

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1977
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Cited by 208 publications
(72 citation statements)
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“…Solution B was the same but without EGTA. These high-K+ media were used to prevent the changes in cell volume, pH and ion content which occur when red cells are permeabilized to divalent cations and K+ in low-K+ media, as when divalent cation ionophores are used in the presence of Ca2+ (Ferreira & Lew, 1976;Lew & Garcia-Sancho, 1989). The washed cells were resuspended at about 10 % packed cell volume (haematocrit; Hct) in solution B with added inosine (10 mM) and "5Ca2" (NEN Research Products, Boston, MA, USA) to give the indicated final calcium concentrations.…”
Section: Methodsmentioning
confidence: 99%
See 1 more Smart Citation
“…Solution B was the same but without EGTA. These high-K+ media were used to prevent the changes in cell volume, pH and ion content which occur when red cells are permeabilized to divalent cations and K+ in low-K+ media, as when divalent cation ionophores are used in the presence of Ca2+ (Ferreira & Lew, 1976;Lew & Garcia-Sancho, 1989). The washed cells were resuspended at about 10 % packed cell volume (haematocrit; Hct) in solution B with added inosine (10 mM) and "5Ca2" (NEN Research Products, Boston, MA, USA) to give the indicated final calcium concentrations.…”
Section: Methodsmentioning
confidence: 99%
“…Several 50 j1 samples were taken before and after ionophore and Co2+ additions for determination of [CaTA as previously described (Dagher & Lew, 1988;Lew & Garcia-Sancho, 1989). Effect of deoxygenation on cytoplasmic Ca2' buffering Cytoplasmic Ca2+ buffering was measured by the method of Ferreira & Lew (1976) (Lew & Garcia-Sancho, 1989) in jnosine-fed and ATP-depleted red cells. ATP was depleted by preincubating the cells at 37 0C for 90 min in medium A with 10 mm inosine and 6 mm iodoacetamide (Lew, 1971).…”
Section: Methodsmentioning
confidence: 99%
“…2B) (Ferreira & Lew, 1976;Simons, 1982 The consistent similarity of Ca2+ desaturation patterns from cells with and without Co2+, illustrated in Fig. 3, emphasises in finer detail than previously reported (Tiffert et al 1984) the lack of any detectable effects of internal Co2+ on Ca2+ pump-mediated Ca2+ extrusion from intact cells.…”
Section: Resultsmentioning
confidence: 68%
“…In such media, red cells permeabilized to divalent cations with the ionophore A23187, and indirectly to K+ by increased [Ca2+]i, maintain their normal volume, pH, rnembrane potential and ionic composition, except for the intended divalent cation concentrationi changes (Ferreira & Lew, 1976: Lew & Brown, 1979Lew & GarciaSancho, 1989). …”
Section: Methodsmentioning
confidence: 99%
“…A common shortcoming of these studies is that the ATP-, Mg2'-and Ca2'-binding 'active centre' of the transport system is in the cell interior and thus adjusting the experimental parameters at this site is inherently difficult. A further problem is the intracellular binding of ATP, Mg" and Ca2+ to various cellular buffer systems [4,8,9] .…”
Section: Introductionmentioning
confidence: 99%