TAGZyme Qcyclase/pGAPase Enzymes (150 U)

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Publication protocol

EnzymaticremovalofN-terminalpyrrolidone carboxylic acid: Solution of pGAPase (25 mL, 25 U/mL) (from the TAG- ZymeTM kit, Qiagen, Hilden, Germany) was activated for 10 min at 377C by adding 10 mL of 200 mM sodium phos- phate (pH 7.5), 10 mL of 800 mM NaCl and 10 mL of freshly prepared 50 mM cysteamine-HCl. Solution of (25 mL, 50 U/ mL) Qcyclase (TAGZyme kit) were then added to the pGA- Pase solution. The dried flow-through of the AccuBONDII SCX SPE cartridge was redissolved in 30 mL of 0.5 M NaOH (the final pH was adjusted to 7.5), the pGAPase/Qcyclase mixture was added and the enzymatic reaction was con tinued for 1 h at 377C.

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