Rapid™ PNGase F (non-reducing format)

The following reagents are supplied with this product:

NEB #	Component Name	Component #	Stored at (°C)	Amount	Concentration
P0711S     4       Rapid™ PNGase F (non-reducing format) P0711SVIAL 4 1 x 0.05 ml 50 reactions   Rapid PNGase F (non-reducing format) Buffer B0717SVIAL 4 1 x 0.2 ml 5 X

Application Features

Rapid PNGase F (non-reducing format) is effective on a variety of antibodies despite their structural diversity. Intact protein characterization using Rapid PNGase F (non-reducing format) is convenient, and complete (reducing biased results). The fast processing time decreases the risk of artifacts and unwanted protein modifications (oxidation, deamidation) that could be introduced by lengthy incubations with standard N-glycan removal protocols. Samples are ready to be prepared for downstream chromatography or mass spectrometry analysis, providing accurate and reproducible results.

For streamlined proteomic workflows, Rapid PNGase F (non-reducing format) is directly compatible with Trypsin-ultra™, Mass Spectrometry Grade (NEB #P8101 ). 

Companion Products

• Rapid PNGase F Antibody Standard
• Endo S
• Trypsin-ultra™, Mass Spectrometry Grade
• Rapid PNGase F

1. Storage: It is recommended to store this kit at 4°C. All components will be stable for at least one year if stored correctly. Do not freeze.
   
2. The pre-incubation step at 75°C is strictly required. Deglycosylationtakes place once the enzyme is added and incubated at 50°C.
3. The target protein should be in a solution compatible with Rapid PNGase F(non-reducing format) activity. Avoid buffers containing SDS, as itinhibits PNGase F. Common stabilizing reagents such as Tween, TritonX-100, NP-40, octyl glucoside and non-detergent sulfobetaine, as well astraces of organic solvents, can prevent optimal rapid deglycosylation.
4. Buffer exchange (by dialysis, spin columns, or microfiltration) isrecommended to prepare samples for liquid chromatography/massspectrometry.

1. Maly, F. et al. (1989). Anal. Biochem. 180, 195-204.

1. Rapid PNGase F (non-reducing format) (P0711) Reaction Protocol
2. Rapid PNGase F (non-reducing format) (P0711) SDS-PAGE Protocol
3. Glycoproteomics: Buffer Exchange Protocols (P0711)

• 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

• Endoglycosidase Selection Chart

1. What is the difference between Rapid PNGase F and Rapid PNGase F (non-reducing format)?
2. What is the difference between Rapid PNGase F (non-reducing format) and Endo S (Endo S, NEB# P0741)?
3. What are the components of Rapid PNGase F (non-reducing format)?
4. How much antibody sample can be deglycosylated with Rapid PNGase F (non-reducing format)?
5. What is a good control substrate for Rapid PNGase F?
6. What should I do if my target protein is only partially deglycosylated with Rapid PNGase F (non-reducing format)?
7. What buffers or additives should be avoided when using Rapid PNGase F (non-reducing format)?
8. How can I test if the Rapid PNGase F (non-reducing format) reaction is complete in 10 minutes?
9. Is Rapid PNGase F (non-reducing format) compatible with downstream mass spectrometry analysis?
10. Is Rapid PNGase F (non-reducing format) only suitable for deglycosylation of antibodies?
11. Are longer incubation times detrimental to the Rapid PNGase F (non-reducing format) reaction?
12. What is the reaction temperature range for Rapid PNGase F (non-reducing format)?
13. What happens to the asparagine residue after Rapid PNGase F removes the sugar?
14. Can a protease inhibitor cocktail be used in a Rapid PNGase F (non-reducing format) reaction?
15. Is the Rapid PNGase F (non-reducing format) reaction compatible with Trypsin-ultra™ (NEB# P8101)?

• Glycoproteomics Brochure
• Glycoproteomics Technical Guide

• Using Glycosidases to Remove Trim or Modify Glycans on Therapeutic Protein
• Glycomics: A rapidly evolving field with a sweet future
• Redesigning glycosidase manufacturing quality for pharmaceutical and clinical applications

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.

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]

• P0711S_v2

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]

• P0711S_v1_0021602
• P0711S_v1_0021605
• P0711S_v1_0021610
• P0711S_v1_0021702
• P0711S_v1_0021704
• P0711S_v1_0021709
• P0711S_v2_10024950
• P0711S_v2_10037455
• P0711S_v2_10047786
• P0711S_v0_10052559
• P0711S_v2_10055358
• P0711S_v2_10057730
• P0711S_v2_10060517
• P0711S_v2_10065636
• P0711S_v2_10075418
• P0711S_v2_10085473
• P0711S_v2_10101281
• P0711S_v2_10106635
• P0711S_v2_10118376
• P0711S_v2_10124414
• P0711S_v2_10146869
• P0711S_v2_10163671
• P0711S_v2_10179236
• P0711S_v2_10185488
• P0711S_v2_10200634
• P0711S_v2_10208875
• P0711S_v2_0021801
• P0711S_v2_10227063
• P0711S_v2_10243892
• P0711S_v2_10253393

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

• Rapid™ PNGase F (non-reducing format)
• Rapid PNGase F (non-reducing format) Buffer

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.

Trademarks

NEW ENGLAND BIOLABS^® is a registered trademark of New England Biolabs, Inc.
RAPID^™ and ULTRA^™ are trademarks of New England Biolabs, Inc.