HpGroEL variants were diluted into the ATPase reaction buffer (50 mM TrisHCl, 1 mM KCl/MgCl 2 , 250 M ATP, pH 7.5) to a final concentration of 1 M
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For analysis of the effect of cell culture conditions, hES-MSCs were seeded at densities of 5 10 3 and 2 10 3 cells/cm 2 , incubated for 19 h, stained with FreSHtracer, and the fluorescence ratio (F 510 /F 580 ) was measured as described above
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The water appears to be positioned to pull off the proton from the sulphur atom of GSH