Selected article for: "additional sample and loading buffer"

Author: Benharouga, Mohamed; Haardt, Martin; Kartner, Norbert; Lukacs, Gergely L.
Title: Cooh-Terminal Truncations Promote Proteasome-Dependent Degradation of Mature Cystic Fibrosis Transmembrane Conductance Regulator from Post-Golgi Compartments
  • Document date: 2001_5_28
  • ID: q3agdeju_23
    Snippet: The plasma membrane cAMP-dependent halide conductance of BHK cells expressing T70 CFTR was determined with iodide efflux as described (Mohamed et al., 1997) . Iodide efflux was initiated by replacing the loading buffer with efflux medium (composed of 136 mM nitrate in place of iodide). The extracellular medium was replaced every minute with efflux buffer (1 ml). After a steady state was reached, the intracellular cAMP level was raised by agonists.....
    Document: The plasma membrane cAMP-dependent halide conductance of BHK cells expressing T70 CFTR was determined with iodide efflux as described (Mohamed et al., 1997) . Iodide efflux was initiated by replacing the loading buffer with efflux medium (composed of 136 mM nitrate in place of iodide). The extracellular medium was replaced every minute with efflux buffer (1 ml). After a steady state was reached, the intracellular cAMP level was raised by agonists (10 M forskolin, 0.2 mM CTP-cAMP, and 0.2 mM isobutyl-methyl xanthane) to achieve maximal phosphorylation of the T70 CFTR. The collection of the efflux medium resumed for an additional 6-9 min. The amount of iodide in each sample was determined with an iodide-selective electrode (Orion).

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