Enzymatic deglycosylation of the 25/45 fraction isolated from 20-day-old rats. The control glycoprotein bovine fetuin (1 μg/lane) and the 25/45 fraction (8 μg/lane) were incubated with (+) or without (-) deglycosylation enzymes (Enz) prior to their electrophoresis through a 5-20% gradient gel and transfer to PVDF. Deglycosylation of fetuin resulted in a major shift in its molecular weight (compare lanes 1 and 2). Minor differences in Coomassie Blue staining patterns are evident between the untreated (lane 3) and deglycosylated (lane 4) 25/45 fraction samples. However, deglycosylation of the 25/45 fraction failed to alter the electrophoretic mobility or immunoreactivity of the antigen recognized by MAb 10A5 (compare lanes 6 and 7).