Yes, but not as efficiently as it will in the DNase I-XT Reaction Buffer (NEB #B0570). DNase I-XT is a calcium-dependent enzyme that works best in DNase I-XT Reaction Buffer, which contains calcium chloride. It is recommended that even if you are using NEB buffers 1-4 you also add DNase I-XT buffer or 0.5 mM CaCl2 to the reaction.