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Next, the At LDOX protein was removed using an Amicon Ultra-4 device (10 kDa cut-off) (Merck Millipore) and the eluent was diluted with 1.375 vol of the same reaction buffer containing 82% H 2 18 O and incubated at 30 C, 1,200 rpm using a 3 mm shaking diameter to achieve 18 O equilibration