PNGase F

PNGase F cleaves N-linked (asparagine-linked) oligosaccharides from glycoproteins.

Enzyme remains fully active for at least 96 hours at 37˚C.

View product documentation:

Part Number – Amount of Enzyme
E-PNG01         – 60 µLs¹
E-PNG01-20   –  20 µLs¹
E-PNG01-200 – 200 µls² (previously E-PNG05)
¹ includes buffer, denaturant, and Triton-X
² includes enzyme only

Product Description


PNGase F, Peptide N Glycosidase F, N-Glycosidase, N-Glycanase

PNGase F, also known as Peptide N-Glycosidase F, is an enzyme commonly used in molecular biology and biochemistry research. It plays a crucial role in the study of glycoproteins, which are proteins that have sugar molecules (glycans) attached to them. PNGase F is used to remove N-linked glycans from glycoproteins, making it a valuable tool in various scientific applications.

Usage of PNGase F:

Glycoprotein Deglycosylation: PNGase F enzymatically cleaves the N-linked glycan chains from glycoproteins. N-linked glycans are a specific type of sugar chain that attaches to asparagine (N) residues in the protein’s amino acid sequence.

Importance in Glycoprotein Analysis: Removing the glycans from glycoproteins is essential for studying the underlying protein structure and function. It allows researchers to analyze the protein core, determine its molecular weight accurately, and investigate its biological activities.

Applications: PNGase F is widely used in various applications, including proteomics, structural biology, immunology, and pharmaceutical research. It is instrumental in characterizing glycoproteins, understanding their roles in cellular processes, and developing therapeutic drugs.

Biomedical Research: In biomedical research, PNGase F is used to analyze glycoproteins associated with diseases such as cancer, diabetes, and infectious diseases. It aids in identifying biomarkers and potential drug targets.

Quality Control: In the biopharmaceutical industry, PNGase F is employed to ensure the quality and consistency of glycoprotein-based drugs, such as monoclonal antibodies, by verifying glycan structures and monitoring glycosylation patterns.

Glycoprotein Engineering: Researchers can use PNGase F to modify glycoproteins for specific purposes, such as improving drug efficacy or altering glycan structures for research or therapeutic applications.

PNGase F Specifications and Protocols:

PNGase F is a versatile enzyme used in the study of glycoproteins, allowing researchers to investigate their structure, function, and significance in various biological processes and applications. Its ability to remove N-linked glycans from glycoproteins makes it a valuable tool in molecular biology and biochemistry research.

PNGase F cleaves N-linked (asparagine-linked) oligosaccharides from glycoproteins. The enzyme deaminates asparagine to aspartic acid, leaving the oligosaccharides intact. PNGase F will not remove oligosaccharides containing Alpha-(1,3)-linked core fucose commonly found on plant glycoproteins; for this purpose, use peptide N-glycosidase A.

Denaturation increases the rate of cleavage. Most native proteins can still be completely N-deglycosylated but incubation time may need to be increased. The enzyme will remain fully active under reaction conditions (37˚C) for at least 96 hours.

There are a number of alternative enzymes which can be used to remove N-glycans, most especially the Endo F family of enzymes and Endo H. These enzymes cleave between the two N-acetylglucosamine residues in the core of the oligosaccharide, generating a truncated sugar molecule with one N-acetylglucosamine residue remaining on the asparagine. This enhances the solubility, keeping proteins in solution that precipitate after deglycosylation with PNGase F which removes the oligosaccharide intact. Endo F1 cleaves high mannose and some hybrid type N-glycans. Endo F2 will removes biantennary and high mannose (at a 40X reduced rate). Endo F3 releases of triantennarry and fucosylated biantennary N-glycans. Endo H removes hybrid or high mannose glycans.

endoglycosidases image
Activity comparison of Endo F1, Endo F2, Endo F3, Endo H, and PNgase F


PNGase F Source Elizabethkingia miricola (was Chryseobacterium meningosepticum)


Contents PNGase F in 20 mM Tris-HCl, pH 7.5

Included with 20 µL and 60 µL pack sizes:
5x Reaction Buffer 7.5 – 250 mM sodium phosphate, pH 7.5
Denaturation Solution – 2% SDS, 1 M Beta-mercaptoethanol
Triton X-100 – 15% solution

Specific Activity >25 U/mg

Activity 5 U/ml

Molecular weight 35,000 daltons

pH range 6-10, optimum 7.5

PNGase F Protocol
1. Add up to 200 µg of glycoprotein to an Eppendorf tube. Adjust to 35 µl final volume with de-ionized water.
2. Add 10 µl 5x Reaction Buffer 7.5 and 2.5 µl of Denaturation Solution. Heat at 100°C for 5 minutes.
3. Cool. Add 2.5 µl of Triton X-100 and mix.
4. Add 2.0 µl of enzyme to the reaction. Incubate 3 hours at 37°C.

Specifictity Cleaves all asparagine-linked complex, hybrid or high mannose oligosaccharides unless alpha(1-3) core fucosylated; asparagine must be peptide bonded at both termini, Endo F free

Specific Activity Defined as the amount of enzyme required to catalyze the release of N-linked oligosaccharides from 1 micromole of denatured RNase B in 1 minute at 37°C, pH 7.5. Cleavage is monitored by SDS-PAGE (cleaved RNase B migrates faster).

Storage Store enzyme at 4°C.


– Bayer, E.A., F. De Meester, T. Kulik and M. Wilchek. Preparation of deglycosylated egg white avidin. Appl Biochem Biotech 53: 1-9 (1995)

– Elder, J.H. and S. Alexander. Endo-b-N-Acetylglucosaminidase F: endoglycosidase from Flavobacterium meningosepticum that cleaves both high-mannose and complex glycoproteins. Proc Natl Acad Sci USA 79: 4540-4544 (1982)

– Tarentino, A .L., C.M. Gomez and T.H. Plummer, Jr. Deglycosylation of asparagine-linked glycans by peptide :N-glycosidase F. Biochemistry 24: 4665-4671 (1985)

– Tarentino A.L. and T.H. Plummer. Enzymatic deglycosylation of asparagine -linked glycans: purification, properties, and specificity of oligosaccharide-cleaving enzymes from Flavobacterium meningosepticum. Meth Enzymol 230: 44-57 (1994)

– Trimble R.B. and A.L. Tarentino. Identification of distinct endoglycosidase (endo) activities in Flavobacterium meningosepticum: endo F1 , endo F2 and endo F3. Endo F1 and endo H hydrolyze only high mannose and hybrid glycans. J Biol Chem 266: 1646-1 651 (1991)

– Taga, E. M., A. Waheed and R. L. Van Etten. Structural and chemical characterization of a homogeneous peptide-N-glycosidase from almond. Biochemistry 23: 815-22 (1984)

– Tarentino AL, Trimble RB, Plummer TH. Enzymatic approaches for studying the structure, synthesis, and processing of glycoproteins. Methods in Cell Biology: 32: 111–39 (1989)

Anthony L. , Tarentino and Thomas H. Plummer Jr. Enzymatic deglycosylation of asparagine-linked glycans: Purification, properties, and specificity of oligosaccharide-cleaving enzymes from Flavobacterium meningosepticum. Methods in Enzymology: 230: 44-57. (1994)

– Tarentino AL, Plummer TH. Oligosaccharide accessibility to peptide:N-glycosidase as promoted by protein-unfolding reagents. The Journal of Biological Chemistry. 257 (18): 10776–80. (1982)