Cy5-Neu5Ac (N-glycan) Labeled Recombinant Human ACE-2

Cy5-Neu5Ac (N-glycan) Labeled Recombinant Human ACE-2 (Cy5-ACE-2) SDS-PAGE. 1 μg/lane of Cy5-ACE-2 was resolved with SDS-PAGE under reducing conditions and visualized by fluorescent imaging (left) and Coomassie blue staining (right), showing a band around 95 kDa. ​

N-glycans from Cy5-Neu5Ac Labeled ACE-2 (Cy5-ACE-2) were released by PNGase F digestion. N-glycans of Cy5-ACE-2 were released by PNGase F N-glycan Releasing Kit (EA006) under denaturing conditions. Cy5-ACE-2 (1 µg per digestion) was treated by PNGase F in 20 µL under at 37 °C for 60 minutes then separated on 17% SDS gel and imaged by trichloroethanol (TCE) staining (top panel) and fluorescent imaging (lower panel). Ten-fold serial dilution of PNGase F was performed starting from 1000 ng from right to left.