NEB, P0704S, PNGase F
PNGase F is the most effective enzymatic method for removing almost all
Catalog Number: P0704S
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| <b>Unit Definition</b> One unit is defined as the amount of enzyme required to remove > 95% of the carbohydrate from 10 µg of denatured RNase B in 1 hour at 37°C in a total reaction volume of 10 μl. Unit Definition Assay | 10 µg of RNase B are denatured with 1X Glycoprotein Denaturing Buffer (0.5% SDS, 40 mM DTT) at 100°C for 10 minutes. After the addition of NP-40 and GlycoBuffer 2, two-fold dilutions of PNGase F are added and the reaction mix is incubated for 1 hour at 37°C. Separation of reaction products are visualized by SDS-PAGE. 1X Glycoprotein Denaturing Buffer 0.5% SDS 40 mM DTT 1X NP-40 1% NP-40 in MilliQ-H 2 O <b>Reaction Conditions</b> 1X GlycoBuffer 2 Incubate at 37°C 1X GlycoBuffer 2 50 mM Sodium Phosphate (pH 7.5 @ 25°C) <b>Storage Buffer</b> 20 mM Tris-HCl 50 mM NaCl 5 mM EDTA 50% Glycerol pH 7.5 @ 25°C <b>Heat Inactivation</b> 75°C for 10 minutes <b>Molecular Weight</b> Apparent: 36000 daltons |
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