RNase B Deglycosylation Protocol (P7817)

  1. In a 10 µl reaction, add 2 µl of RNase B, 1 µl of 10X Glycoprotein Denaturing Buffer and 5 µl of dH2O.
  2. Incubate at 100°C for 10 minutes.
  3. Add 1 µl of 10X GlycoBuffer 2 and 1 µl of 10% NP-40.
  4. Add 1 µl of endoglycosidase.
  5. Incubate at 37°C for 1 hour.
  6. Visualize reaction products on 10–20% SDS-PAGE gel. 

Note: 250 µg is enough for approximately 20 reactions.