PNGase F

GeneTex

PNGase F

9

One unit is defined as the amount of enzyme required to remove > 95% of the carbohydrate from 10 ug of denatured RNase B in 1 hour at 37°C in a total reaction volume of 10 ul. Protocols for both denaturing and non-denaturing conditions: Reaction buffer (not included; must be prepared by user): Buffer 1 (5% SDS, 0.4 M DTT) Buffer 2 (0.5 M Sodium Phosphate, pH 7.5) 10% IGEPAL® CA-630 Denaturing conditions: Combine 10-20 ug of glycoprotein, 1 ul Buffer 1 (5% SDS, 0.4 M DTT), and H2O to a total volume of 10 ul. Denature glycoprotein by heating at 100°C for 10 minutes. Cool glycoprotein on ice. Add 2 ul Buffer 2 (0.5 M Sodium Phosphate, pH 7.5), 2 ul 10% IGEPAL® CA-630, 2 ul PNGase F and H2O to a total volume of 20 ul. Incubate at 37°C for 1 hour. Analyze by SDS-PAGE. Non-denaturing conditions: Combine 10-20 ug of glycoprotein, 1 ul Buffer 2 (0.5 M Sodium Phosphate, pH 7.5), 1 ul PNGase F, and H2O to a total volume of 10 ul. Incubate at 37°C for 16-20 hours. Analyze by SDS-PAGE.

Research Use Only

500 units/ul

Unconjugated

2°C to 8°C

12352202