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based on consistent formation of -helical structure in initial rounds of structure calculations in which no hydrogen bond restraints were defined), inter-molecular PRE restraints (4 non-interacting copies of the spin label to account for different spin label orientations), inter- and intra-molecular NOE restraints, and ambiguous distance restraints for residues showing CSPs upon binding

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Cells were washed with PBS, dissociated in 10 mM EDTA (2 ml per well) at 37 C for 5 min, and washed three times in cold PBS

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- Multiple Use: Unlike regular sterile water (which is single-use and must be discarded after opening), bacteriostatic water can be used multiple times within a 28-day period.