3.7. Af-EGL7 Deglycosylation by Endo H

Purified Af-EGL7 deglycosylation was accomplished by Endoglycosidase H (Endo H, New England Biolabs, Ipswich, MA, USA) in both denaturing and non-denaturing conditions, according to the manufacturer’s instructions. In the first case, the recombinant protein was initially denatured at 100 °C for 10 min. Then, the deglycosylation reaction was performed at 37 °C for 24 h, as described by Meleiro et al. (2017) [34]. Proteins were further analyzed by SDS-PAGE.