PNGase F (Glycerol-free), Recombinant

Catalog #	Concentration	Size	List Price	Quantity	Your Price
P0709L	500000 units/ml	75000 units	$1,072.00		$964.80
P0709S	500000 units/ml	15000 units	$267.00		$240.30

Catalog #	Size	List Price	Your Price
P0709L	75000 units	$1,072.00	$964.80
P0709S	15000 units	$267.00	$240.30

Peptide: N-Glycosidase F, also known as PNGase F, is a recombinant amidase which is supplied glycerol free for optimal performance in HPLC intensive methods. PNGase F cleaves between the innermost GlcNAc and asparagine residues of high mannose, hybrid, and complex oligosaccharides from N-linked glycoproteins (1).

Product Source

Cloned from Elizabethkingia miricola (formerly Flavobacterium meningosepticum) and expressed in E. coli (2).

Reagents Supplied

The following reagents are supplied with this product:

NEB #

Component Name

Component #

Stored at (°C)

Amount

Concentration

P0709S 4

PNGase F (Glycerol-free), Recombinant	P0709SVIAL	4	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 %

P0709L 4

PNGase F (Glycerol-free), Recombinant	P0709LVIAL	4	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

Properties & Usage

Unit Definition

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.

1X Glycoprotein Denaturing Buffer
0.5% SDS
40 mM DTT

1X NP-40
1% NP-40 in MilliQ-H₂O

Reaction Conditions

1X GlycoBuffer 2
Incubate at 37°C

1X GlycoBuffer 2
50 mM Sodium Phosphate
(pH 7.5 @ 25°C)

Storage Buffer

50 mM NaCl
20 mM Tris-HCl
5 mM EDTA
pH 7.5 @ 25°C

Heat Inactivation

75°C for 10 minutes

Molecular Weight

Apparent: 36000 daltons

Unit Assay Conditions

10 μg of RNase B are denatured with 1X Glycoprotein Denaturing Buffer at 100°C for 10 minutes. After the addition of NP-40 and GlycoBuffer 2, two-fold dilutions of PNGase F (Glycerol-free), Recombinant are added and the reaction mix is incubated for 1 hour at 37°C. Separation of reaction products are visualized by SDS-PAGE.

Companion Products

Notes

To deglycosylate a native glycoprotein, longer incubation time as well as more enzyme may be required.
Since PNGase F (Glycerol-free), 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.
PNGase F (Glycerol-free), Recombinant will not cleave N-linked glycans containing core α1-3 Fucose.
Recommended storage temperature is 4°C, avoid repeat freeze-thaw cycles.

References

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Chen, M. New England Biolabs, Inc., Unpublished observation

Protocols

PNGase F Protocol

Usage Guidelines

Selection Charts

Endoglycosidase Selection Chart

FAQs

Tech Tips

1. You can use this enzyme under native or denaturing conditions
2. Under native conditions we recommend adding more enzyme and using longer incubation times
3. PNGase F activity is inhibited by SDS, therefore under denaturing conditions it is essential to have NP-40 present in the reaction mixture in a 1:1 ratio.
4. PNGase F will not cleave N-linked glycans containing core α1-3 Fucose (PNGase A must be used in this instance)
5. A good positive control substrate is RNase B

Citations

See more details on Bioz

Quality Control Assay

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

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]

P0709S_L_v1

Certificate of Analysis

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]

Safety Data Sheets

Legal And Disclaimer

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.