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Mouse islets were subsequently washed, purified using a Histopaque gradient (Histopaque-1119 and Histopaque-1083, Sigma-Aldrich) and rested overnight at 37C in a 95% O 2 / 5% CO 2 incubator before performing experiments in mouse islet culture medium (RPMI-1640 with 11 mM D-glucose, supplemented with 10% FBS and 1% penicillin/streptomycin)

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It has been observed that the active-site Ca 2+ coordinates with four acidic residues and if any of the residues are mutated then a greater Ca 2+ concentration is needed for catalysis

Wilding JPH, Batterham RL, Davies M, Van Gaal LF, Kandler K, Konakli K, Lingvay I, McGowan BM, Oral TK, Rosenstock J, Wadden TA, Wharton S, Yokote K, Kushner RF

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