When I analyze my secreted protein by SDS-PAGE and/or Western analysis there is a discrepancy between the predicted and the observed mass. What could cause this?
The most likely cause for a discrepancy between the predicted and observed mass of a secreted protein is the presence of a covalent modification like an N-linked oligosaccharide. Treatment of proteins with endo-glycosidases that remove N-linked sugars (Endo-H #P0702S or PNGase F #P0704S) can help diagnose if the secreted protein is glycosylated.
Related Questions
- When I analyze my secreted protein by SDS-PAGE and/or Western analysis there is a discrepancy between the predicted and the observed mass. What could cause this?
- Can three-dimensional contacts in protein structures be predicted by analysis of correlated mutations?
- Can a protein be identified by mass spec if it is detectable by western blot?
