Remove-iT^® PNGase F

The following reagents are supplied with this product:

NEB #	Component Name	Component #	Stored at (°C)	Amount	Concentration
P0706S     4       Remove-iT® PNGase F P0706SVIAL 4 1 x 0.03 ml 225,000 units/ml   GlycoBuffer 2 B3704SVIAL -20 1 x 1 ml 10 X   DTT B0706SVIAL -20 1 x 1 ml 0.4 M
P0706L     4       Remove-iT® PNGase F P0706LVIAL 4 1 x 0.15 ml 225,000 units/ml   GlycoBuffer 2 B3704SVIAL -20 1 x 1 ml 10 X   DTT B0706SVIAL -20 1 x 1 ml 0.4 M

Companion Products

• RNase B
• Chitin Magnetic Beads
• 6-Tube Magnetic Separation Rack
• 12-Tube Magnetic Separation Rack
• Fetuin
• Endo S
• Endo D
• PNGase F

1. To deglycosylate a native glycoprotein, longerincubation time, as well as more enzyme, maybe required.
2. Using typical RNase B denaturing conditions with NEB Glycoprotein Denaturing Buffer,containing SDS and DTT, Remove-iT PNGase F yields a higher concentration of 500,000 U/ml.
3. If using Remove-iT PNGase F under typical PNGase F denaturing conditions, it is essential to have NP-40 in the reaction mixture as Remove-iT PNGase F is inhibited by SDS. It is not known why this non-ionic detergent counteracts the SDS inhibition.
4. Remove-iT PNGase F will not cleave N-linkedglycans containing core α1-3 Fucose.
5. Recommended storage temperature is 4°C,avoid repeat freeze-thaw cycles
6. Removal of Remove-iT PNGase F from the deglycosylation reaction can be scaled up linearly with larger volumes of chitin magnetic beads.
7. Chitin Magnetic Beads Binding Capacity is0.4 µg/µl of CBD-tagged protein.
8. To remove 1-5 µl of Remove-iT PNGase F use 50 µl of chitin magnetic beads.

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

1. Remove-iT® PNGase F Magnetic Chitin Bead Protocol (P0706)
2. Reaction Conditions for Remove-iT® PNGase F (P0706)

• AppNote_Remove-iT_PNGase_F_Effective_Release_and_Recovery_of_Neutral_and_Sialylated_N-glycans
• 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
• Remove iT PNGase F Effective Release and Recovery of Neutral and Sialylated N glycans

• Endoglycosidase Selection Chart

1. What is the tag on PNGase F?
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 is the difference between PNGase F and Remove-iT® PNGase F?
4. What is the difference between Remove-iT® PNGase F and Endo H?
5. How much Remove-iT® PNGase F should I use to remove my carbohydrate under native or DTT denaturing conditions?
6. Does Remove-iT® PNGase F work in Urea?
7. What are the typical reaction conditions for Remove-iT® PNGase F?
8. How do I eliminate Remove-iT® PNGase F from a reaction?
9. What is the binding capacity of the Magnetic Chitin Beads used to eliminate Remove-iT® PNGase F?
10. Is Remove-iT® PNGase F compatible with downstream analysis such as HPLC and Mass Spectrometry?
11. 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?
12. What is a good endoglycosidase substrate?
13. Do detergents inhibit exoglycosidases/endoglycosidases?
14. What are Glycosidases and their uses?

• Glycoproteomics Brochure
• Glycoproteomics Technical Guide
• Purification Beads, Columns and Resins Brochure

• Using Glycosidases to Remove Trim or Modify Glycans on Therapeutic Protein
• 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.

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]

• P0706S_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]

• P0706S_L_v1_0031607
• P0706S_L_v1_0031701
• P0706S_L_v1_0031707
• P0706S_L_v1_0031712
• P0706S_L_v1_0031805
• P0706S_v1_10018033
• P0706L_v1_10018032
• P0706L_v1_10033433
• P0706S_v1_10027209
• P0706L_v1_10034629
• P0706S_v1_10034628
• P0706L_v1_10042345
• P0706S_v1_10042343
• P0706L_v1_10052760
• P0706S_v1_10052763
• P0706L_v1_10058023
• P0706L_v1_10062846
• P0706S_v1_10058024
• P0706L_v1_10067111
• P0706S_v1_10062848
• P0706S_v1_10077466
• P0706S_v1_10081120
• P0706L_v1_10081122
• P0706S_v1_10092463
• P0706L_v1_10098748
• P0706S_v1_10102192
• P0706L_v1_10102195
• P0706S_v1_10123406
• P0706L_v1_10126945
• P0706L_v1_10153983
• P0706S_v1_10153981
• P0706L_v1_10179182
• P0706S_v1_10182792
• P0706S_v1_10213125
• P0706L_v1_10217282
• P0706S_v1_10238177
• P0706S_v1_10238461
• P0706L_v1_10238178
• P0706S_v1_10245387
• P0706L_v1_10256171

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

• Remove-iT^® PNGase F
• GlycoBuffer 2
• DTT

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.
REMOVE-IT™ is a trademark of New England Biolabs, Inc.