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In the second step, conjugating enzymes perform addition reactions (e.g., acetyl, methyl, glucuronic acid) on these newly modified, electrophilic substrates

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Recombinant proteins were analyzed using 10% SDS-PAGE and purified using HisTrap HP Crude Columns (GE Health) and AmiconUltra-15 Centrifugal Filter Units with Ultracel-3 membranes (Merck Millipore)

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