PNGase F

Species: Elizabethkingia miricola

Product Description: Peptide:N-glycosidase F (PNGase F) removes all N-linked glycans (high-mannose, hybrid, and complex) from glycopeptides and glycoproteins. This PNGase F, a recombinant amidase cloned from Elizabethkingia miricola, cleaves between the innermost GlcNAc and asparagine residues. Protocols for both denaturing and non-denaturing conditions are shown in the "Application Note" section.

One vial of GTX141281-pro contains 75000 units (75kU) PNGase F.

Application Note:

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.

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:

1. Combine 10-20 µg of glycoprotein, 1 µl Buffer 1 (5% SDS, 0.4 M DTT), and H₂O to a total volume of 10 µl.
2. Denature glycoprotein by heating at 100°C for 10 minutes.
3. Cool glycoprotein on ice.
4. Add 2 µl Buffer 2 (0.5 M Sodium Phosphate, pH 7.5), 2 µl 10% IGEPAL® CA-630, 2 µl PNGase F and H₂O to a total volume of 20 µl.
5. Incubate at 37°C for 1 hour.
6. Analyze by SDS-PAGE.

Non-denaturing conditions:

1. Combine 10-20 µg of glycoprotein, 1 µl Buffer 2 (0.5 M Sodium Phosphate, pH 7.5), 1 µl PNGase F, and H₂O to a total volume of 10 µl.
2. Incubate at 37°C for 16-20 hours.
3. Analyze by SDS-PAGE.

Form: Liquid

Buffer (with preservative): PBS, 10% Glycerol, 5 mM EDTA, no Preservative.

Concentration: 500 Unit/μl (Please refer to the vial label for the specific concentration.)

Region Sequence: PNGase F protein (41-354 a.a. of #P21163 )

ExpressionSystem: E. Coli

Conjugation: Unconjugated

• SKU: GTX141281-PRO-75
• Manufacturer: GeneTex
• Manufacturer SKU: GTX141281-PRO-75
• Package Unit: 75 kU
• Quantity Unit: STK
• Reactivity: Human