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The inclusion bodies thus obtained were precipitated using 65% ice-cold acetone
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As for azurin (12), lowering the freezing temperature from -5°C to -13°C increases the number of binding sites but the average affinity is lowered
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Although we predict similar volume changes for this association in the two different environments, the apparent affinities will differ because the effective concentration of membrane-bound proteins tends to be much higher due to their confinement on the reduced area of the membrane surface giving stronger apparent affinities (see Runnels and Scarlata, 1999)
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