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The cell pellet was resuspended in lysis buffer (20 mM HEPES pH 7.5, 300 mM NaCl, 5% glycerol, 0.1% NP-40, 5 mM beta-mercaptoethanol, AEBSF and Serva protease inhibitor cocktail) and sonicated

Limited or no added energy support

[PubMed: 15946163] 171

This is the reason that each subsequent treatment must be spaced out appropriately

Fluid redistribution in sleep apnea: therapeutic implications in edematous states