PNGase F, Recombinant

Reagents Supplied

The following reagents are supplied with this product:

NEB #	Component Name	Component #	Stored at (°C)	Amount	Concentration
P0708S     -20       PNGase F, Recombinant P0708SVIAL -20 1 x 0.03 ml 500,000 units/ml   GlycoBuffer 2 B3704SVIAL -20 1 x 1 ml 10 X   Glycoprotein Denaturing Buffer B1704SVIAL -20 1 x 1 ml 10 X   NP-40 B2704SVIAL -20 1 x 1 ml 10 %
P0708L     -20       PNGase F, Recombinant P0708LVIAL -20 1 x 0.15 ml 500,000 units/ml   GlycoBuffer 2 B3704SVIAL -20 1 x 1 ml 10 X   Glycoprotein Denaturing Buffer B1704SVIAL -20 1 x 1 ml 10 X   NP-40 B2704SVIAL -20 1 x 1 ml 10 %

Application Features

• Removal of carbohydrate residues from proteins

Companion Products

• RNase B
• Endoglycosidase Reaction Buffer Pack
• Endo H
• Endo H_f
• Remove-iT® PNGase F
• O-Glycosidase

1. To deglycosylate a native glycoprotein, longer incubation time as well as more enzyme may be required.
2. Since PNGase F, Recombinant activity is inhibited by SDS, it is essential to have NP-40 in the reaction mixture. It is not known why this non-ionic detergent counteracts the SDS inhibition at the present time.
3. PNGase F, Recombinant will not cleave N-linked glycans containing core α1-3 Fucose.

1. Maley, F. et al. (1989). Anal. Biochem. 180, 195-204.
2. Chen, M. New England Biolabs, Inc., Unpublished observation

1. PNGase F Protocol

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

• AppNote_Remove-iT_PNGase_F_Effective_Release_and_Recovery_of_Neutral_and_Sialylated_N-glycans
• AppNote_Unbiased_and_Fast_IgG_Deglycosylation _for_Accurate_N-glycan_Analysis_using_Rapid_PNGase_F
• AppNote_Proteomics_Fast_and_Efficient_Antibody_Deglycosylation_using_Rapid_PNGase_F
• AppNote_Glycan_Analysis_of_Murine_IgG2a_by_Enzymatic_Digestion_with_PNGase_F_and_Trypsin
• Glycan Analysis of Murine IgG2a by Enzymatic Digestion with PNGase F and Trypsin, Followed by Mass Spectrometric Analysis
• Detailed Characterization of Antibody Glycan Structure using the N-Glycan Sequencing Kit
• 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

• Endoglycosidase Selection Chart

1. Is there a difference between PNGase F (P0704/P0705) and PNGase F, Recombinant (P0708/P0709)?
2. I tried deglycosylating my glycoprotein with Remove-iT® PNGase F but did not see removal of the carbohydrate. What could be the problem?
3. What happens to the asparagine after PNGase removes the sugar?
4. Why is protein degraded? When I denature and add SDS all I see on my SDS-PAGE is a smear or no protein. Can a protease inhibitor cocktail be used in a PNGase F, Recombinant reaction?
5. What is the difference between PNGase F, Endo H and O-Glycosidase?
6. Does PNGase F work in Urea?
7. What are the typical reaction conditions for PNGase F, Recombinant?
8. How much PNGase F, Recombinant should I use to remove my carbohydrate under native or DTT denaturing conditions?
9. How do I inhibit PNGase F, Recombinant?
10. Is PNGase F, Recombinant compatible with downstream analysis such as HPLC and Mass Spectrometry?
11. What are glycosidases and their uses?
12. Do detergents inhibit exoglycosidases/endoglycosidases?
13. 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?
14. What is a good endoglycosidase substrate?

• Glycoproteomics Brochure
• Glycoproteomics Technical Guide

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

Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.

Specifications & Change Notifications

The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]

• P0708S_L_v1

The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]

• P0708S_L_v1_0021605
• P0708S_L_v1_0021611
• P0708S_L_v1_0021709
• P0708S_L_v1_0021711
• P0708S_L_v1_0021801
• P0708S_v1_10007352
• P0708L_v1_10007350
• P0708L_v1_10018428
• P0708S_v1_10024416
• P0708L_v1_10024414
• P0708S_v1_10044573
• P0708L_v1_10044575
• P0708L_v1_10064923
• P0708S_v1_10064924
• P0708S_v1_10088306
• P0708L_v1_10088308
• P0708L_v1_10106551
• P0708S_v1_10106553
• P0708S_v1_10137161
• P0708L_v1_10142232
• P0708S_v1_10156144
• P0708L_v1_10156146
• P0708L_v1_10182244

The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.

• PNGase F, Recombinant
• GlycoBuffer 2
• Glycoprotein Denaturing Buffer
• NP-40

Products and content are covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB). The use of trademark symbols does not necessarily indicate that the name is trademarked in the country where it is being read; it indicates where the content was originally developed. The use of this product may require the buyer to obtain additional third-party intellectual property rights for certain applications. For more information, please email busdev@neb.com.

This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

New England Biolabs (NEB) is committed to practicing ethical science – we believe it is our job as researchers to ask the important questions that when answered will help preserve our quality of life and the world that we live in. However, this research should always be done in safe and ethical manner. Learn more.