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Protein was eluted with 4CV of 60% buffer B and immediately diluted 10-fold with Buffer A prior to concentration and 3 rounds of buffer exchange to remove excess imidazole by centrifugation at 3095xg at 4 C in 100 kDa cutoff concentrators (Amicon)

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10.1016/j.ejpb.2016.11.026 28 CedrowskaE.PruszynskiM.GawedaW.ZukM.KrysinskiP.BruchertseiferF.et al (2020)

Fractions were analyzed by SDS PAGE and those identified to contain tag-free GRXs were combined and subsequently spin-concentrated with a VivaSpin MWCO 3k concentrator at 4000 rpm and 4 C