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The reason its LogD does not decrease appreciably after it is deprotonated is, again, because the negative charge is delocalized within the molecule in internal hydrogen bonding, such that it becomes a lipophilic anion and so can act as the all-important intramembrane proton shunt (28)

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After incubation, wash coated beads once with bulk lysis buffer and twice with wash buffer (50 mM HEPES at pH 7.3, 500 mM KCl) and resuspend beads in 60 l wash buffer