Journal: bioRxiv

Article Title: Systems-wide analysis of glycoprotein conformational changes by limited deglycosylation assay

doi: 10.1101/2021.06.04.447131

Figure Lengend Snippet: Experimental conditions for optimal limited deglycosylation of bovine RNase B and fetuin by PNGase F were determined. The optimal A ) NP-40 concentration, B ) temperature, C ) and D ) PNGase F:glycoprotein ratio for RNase B and fetuin, respectively, and, E ) and F ) the optimal time for the deglycosylation of RNase B and fetuin, respectively, were assayed by differential migration of the glycoproteins in 15% SDS-PAGE. Fully deglycosylated (+) and fully glycosylated (−) proteins were included for each optimization reaction, denoted by FD and FG, respectively.

Article Snippet: Proof-of-concept and optimization of the LDA method were performed using two standard glycoproteins, bovine RNase B and fetuin (New England Biolabs) by PNGase F (New England Biolabs) under native conditions.

Techniques: Concentration Assay, Migration, SDS Page