PNGase F (Glycerol-free)

Application Features

• Removal of N-linked glycans from glycoproteins
• Preferred formulation for HPLC and MS intensive methods of glycoprotein analysis

Companion Products

• Endo H
• Endo H_f
• PNGase F
• Remove-iT® PNGase F
• O-Glycosidase
• Endo S
• Endo D
• RNase B

1. Since PNGase F activity is inhibited by SDS, it is essential to have NP-40 present in the reaction mixture. Why this non-ionic detergent counteracts the SDS inhibition is unknown at present.
2. To deglycosylate a native glycoprotein, longer incubation time as well as more enzyme may be required. 
   
3. PNGase F will not cleave N-linked glycans containing core α1-3 Fucose. 
4. Repeated freeze thaw cycles degrade enzyme activity over time. 
   
5. Typical reaction conditions: Please see Protocol tab

1. PNGase F Protocol

• Detailed Characterization of Several Glycosidase Enzymes
• Glycobiology Unit Conversion Chart

• Characterization of glycans from Erbitux Rituxan and Enbrel using PNGase F Glycerol-free Recombinant
• Remove-iT PNGase F Effective Release and Recovery of Neutral and Sialylated N-glycans
• Enzymatic removal of N- and O-glycans using PNGase F or the Protein Deglycosylation_Mix
• Unbiased_and fast IgG deglycosylation for accurate N-glycan analysis using Rapid PNGase F
• Proteomics_Fast and Efficient Antibody Deglycosylation using Rapid PNGase F
• Glycan Analysis of Murine lgG2a by Enzymatic Digestion with PNGase F and Trypsin Followed by Mass Sp
• Glycan Analysis of Murine IgG2a by Enzymatic Digestion with PNGase F and Trypsin, Followed by Mass Spectrometric Analysis

• Endoglycosidase Selection Chart

1. Is PNGase F compatible with downstream analysis such as HPLC and Mass Spectrometry?
2. Why is my immunoprecipitated (IP) protein degraded? When I denature and add SDS all I see on my SDS-PAGE is a smear or no protein?
3. Do detergents inhibit exoglycosidases/endoglycosidases?
4. Does PNGase F work in Urea?
5. How do I inhibit PNGase F?
6. How much PNGase F should I use to remove my carbohydrate under native conditions?
7. What is the difference between PNGase F, Endo H and O-Glycosidase?
8. I tried the PNGase F on my glycoprotein and didn't see removal of the carbohydrate. What could be the problem?
9. Why have the NEB Glycosidase enzymes changed reaction buffers? What are the new reaction buffers and can I still use an enzyme with its old buffer? Where can I find the composition of the old buffers?
10. What are glycosidases and their uses?

• Glycoproteomics Brochure
• Glycoproteomics Technical Guide

Product Citation Tool

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• N-Glycan Composition Profiling for Quality Testing of Biotherapeutics
• N-Glycan Composition Profiling for Quality Testing of Biotherapeutics
• Redesigning glycosidase manufacturing quality for pharmaceutical and clinical applications
• Using Glycosidases to Remove Trim or Modify Glycans on Therapeutic Protein

• P0705S_L_v1

• P0705S_L_v1_0421507
• P0705S_L_v1_0421603
• P0705S_L_v1_0421512
• P0705S_L_v1_0431609
• P0705S_L_v1_0431705
• P0705S_L_v1_0431712
• P0705S_v1_10013523
• P0705L_v1_10013522
• P0705S_v1_10025789
• P0705L_v1_10027181
• P0705S_v1_10032249
• P0705L_v1_10039782
• P0705S_v1_10039780
• P0705S_v1_10040401
• P0705L_v1_10040400
• P0705L_v1_10045953
• P0705S_v1_10045952
• P0705S_v1_10049548
• P0705S_v1_10052050

• PNGase F (Glycerol-free)
• Glycoprotein Denaturing Buffer
• NP-40
• GlycoBuffer 2