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Basolateral membranes from rat jejunal enterocytes have been obtained by self-orienting Percoll-gradient centrifugation. Bicarbonate and L-glucose uptake into osmotically active basolateral membrane vesicles has been studied by a rapid filtration technique. In closed vessels and at pH 8.2 the uptake kinetics of both [14C]bicarbonate and L[3H]glucose have been followed for 30 min at 18 degrees C. Bicarbonate uptake seems to be fast and in efflux experiments SITS and DIDS effect is negligible. This work demonstrates that it is possible to determine bicarbonate flux across basolateral membrane vesicles at pH and temperature values close to usual experimental conditions.
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