New England Biolabs (NEB) offers a selection of endoglycosidases, exoglycosidases, and heparinases for glycobiology research. Many of these reagents are recombinant, and all undergo several quality control assays, enabling us to provide products with lower unit cost, high purity and reduced lot-to-lot variation. All of our glycosidases are tested for contaminants. Since p-nitrophenyl-glycosides are not hydrolyzed by some exoglycosidases, we use only fluorescently-labeled oligosaccharides to screen for contaminating glycosidases.
To find all of our Glycobiology Application Notes, click here.
- What are glycosidases and their uses?
- Can glycosidases be used in combination for extensive digestion?
- Can I analyze protein glycosylation with only benchtop instrumentation and reagents?
- Do I need to deglycosylate my protein sample for proteomic analysis?
- How can I separate a deglycosylated protein from the glycosidase?
- What are the advantages of enzymatic deglycosylation?
- What are the best techniques to analyze N- and O-glycans?
N-Glycan Composition Profiling for Quality Testing of Biotherapeutics
The Structure, Function and Importance of Carbohydrates
Read about the structure, function, and importance of Carbohydrates from biology experts at NEB.
Using Glycosidases to Remove Trim or Modify Glycans on Therapeutic Protein
- Glycoproteomics Brochure
- Glycoproteomics Technical Guide
- Rapid PNGase F Trifold
- Glycan Analyzer
- Endoglycosidase Selection Chart
- Detailed Characterization of Several Glycosidase Enzymes
- Glycobiology Unit Conversion Chart
- Improving Sample Workflow: Rapid PNGase F for Accurate Antibody Characterization (2015)
- Kalidas, C., Joshi, L., Batt, C. 2001. Characterization of glycosylated variants of beta-lactoglobulin expressed in Pichia pastoris Protein Eng. . 14 , PubMedID: 11342718, DOI:
- Magnelli PE, Bielik AM, Guthrie EP. 2011. Identification and Characterization of Protein Glycosylation using Specific Endo- and Exoglycosidases J Vis Exp. . 58 , PubMedID: 21987255, DOI: 10.3791/3749
- Whistler, J.L., Enquist, J., Marley, A., Fong, J., Gladher, F., Tsuruda, P., Murray, S.R., Von Zastrow, M. 2002. Modulation of postendocytic sorting of G protein-coupled receptors Science . 297 , PubMedID: 12142540, DOI:
- Botto L, Cunati D, Coco S, Sesana S, Bulbarelli A, Biasini E, Colombo L, Negro A, Chiesa R, Masserini M, Palestini P 2014. Role of lipid rafts and GM1 in the segregation and processing of prion protein PLoS One. 9(5), PubMedID: 24859148, DOI: 10.1371/journal.pone.0098344
- Rosenbaum EE, Vasiljevic E, Brehm KS, Colley NJ 2014. Mutations in four glycosyl hydrolases reveal a highly coordinated pathway for rhodopsin biosynthesis and N-glycan trimming in Drosophila melanogaster PLoS Genet. 10(5), PubMedID: 24785692, DOI: 10.1371/journal.pgen.1004349
- Rosenbaek LL, Kortenoeven ML, Aroankins TS, Fenton RA 2014. Phosphorylation decreases ubiquitylation of the thiazide-sensitive cotransporter NCC and subsequent clathrin-mediated endocytosis J Biol Chem. 289(19), PubMedID: 24668812, DOI: 10.1074/jbc.M113.543710
- Möykkynen T, Coleman SK, Semenov A, Keinänen K 2014. The N-terminal domain modulates α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor desensitization J Biol Chem. 289(19), PubMedID: 24652293, DOI: 10.1074/jbc.M113.526301
- Arakel EC, Brandenburg S, Uchida K, Zhang H, Lin YW, Kohl T, Schrul B, Sulkin MS, Efimov IR, Nichols CG, Lehnart SE, Schwappach B 2014. Tuning the electrical properties of the heart by differential trafficking of KATP ion channel complexes J Cell Sci. 127(Pt 9), PubMedID: 24569881, DOI: 10.1242/jcs.141440
- Itahana Y, Han R, Barbier S, Lei Z, Rozen S, Itahana K 2014. The uric acid transporter SLC2A9 is a direct target gene of the tumor suppressor p53 contributing to antioxidant defense Oncogene. , PubMedID: 24858040, DOI: 10.1038/onc.2014.119
- Wicht O, Burkard C, de Haan CA, van Kuppeveld FJ, Rottier PJ, Bosch BJ 2014. Identification and Characterization of a Proteolytically Primed Form of the Murine Coronavirus Spike Proteins after Fusion with the Target Cell J Virol. 88(9), PubMedID: 24554652, DOI: 10.1128/JVI.03451-13
- Wright CR, Brown EL, Della-Gatta PA, Ward AC, Lynch GS, Russell AP 2014. G-CSF does not influence C2C12 myogenesis despite receptor expression in healthy and dystrophic skeletal muscle Front Physiol. 5, PubMedID: 24822049, DOI: 10.3389/fphys.2014.00170
- Zhao H, Blazanovic K, Choi Y, Bailey-Kellogg C, Griswold KE 2014. Gene and protein sequence optimization for high-level production of fully active and aglycosylated lysostaphin in Pichia pastoris Appl Environ Microbiol. 80(9), PubMedID: 24561590, DOI: 10.1128/AEM.03914-13
- Haller G, Li P, Esch C, Hsu S, Goate AM, Steinbach JH 2014. Functional characterization improves associations between rare non-synonymous variants in CHRNB4 and smoking behavior PLoS One. 9(5), PubMedID: 24804708, DOI: 10.1371/journal.pone.0096753
- Stech M, Quast RB, Sachse R, Schulze C, Wüstenhagen DA, Kubick S 2014. A continuous-exchange cell-free protein synthesis system based on extracts from cultured insect cells PLoS One. 9(5), PubMedID: 24804975, DOI: 10.1371/journal.pone.0096635
- Sielaff I. et al. 2006. Protein function microarrays based on self-immobilizing and self-labeling fusion proteins ChemBioChem. . 7 , PubMedID: 16342318, DOI:
- Kwon HM, Lee KH, Han BW, Han MR, Kim DH, Kim DE 2014. An RNA aptamer that specifically binds to the glycosylated hemagglutinin of avian influenza virus and suppresses viral infection in cells PLoS One. 9(5), PubMedID: 24835440, DOI: 10.1371/journal.pone.0097574
- Eichler, R., Lenz, O., Garten, W., Strecker, T. 2006. The role of single N-glycans in proteolytic processing and cell surface transport of the Lassa virus glycoprotein.GP-C. Virol. J. . 3 , PubMedID: 16737539, DOI:
- O’Reilly, M.K., Zhang, G., Imperiali, B. 2006. In vitro evidence for the dual function of Alg2 and Alg11: essential mannosyltransferases in N-linked glycoprotein biosynthesis Biochemistry . 8 , PubMedID: 16878994, DOI:
- Hagen, B.M., Sanders, K.M. 2006. Deglycosylation of the beta1-subunit of the BK channel changes its biophysical properties Am. J. Physiol.Cell. Physiol. . 1 , PubMedID: 16738006, DOI:
- Srikrishna, G., Brive, L., Freeze, H.H. 2005. Novel carboxylated N-glycans contain oligosaccharide-linked glutamic acid Biochem. Biophys. Res. Commun. . 332 , PubMedID: 15922305, DOI:
- Cipollo, J.F., Awad, A.M., Costello, C.E., Hirschberg, C.B. 2005. N-Glycans of Caenorhabditis elegans are specific to developmental stages J. Biol. Chem. . 280 , PubMedID: 15899899, DOI:
- Lüftenegger, D., Picard-Maureau, M., Stanke, N., Rethwilm, A., Lindemann, D. 2005. Analysis and function of prototype foamy virus envelope N glycosylation J. Virol. . 79 , PubMedID: 15919919, DOI:
- Wong-Madden, S.T., Landry, D., and Guthrie, E.P. 1997. Discovery and Uses of Novel Glycosidases Techniques in Glycobiology. , PubMedID: , DOI:
- Magnelli P, Bielik A, Guthrie E. 2012. Identification and characterization of protein glycosylation using specific endo- and exoglycosidases Methods Mol Biol. . 801 , PubMedID: 21987255, DOI:
- Teuton, J.R., Brandt, C.R. 2007. Sialic acid on herpes simplex virus type 1 envelope glycoproteins is required for efficient infection of cells J. Virol. . 81 , PubMedID: 17229687, DOI:
- Song, Y., Choi, M.H., Park, J.N., Kim, M.W., Kim, E.J., Kang, H.A., Kim, J.Y. 2007. Engineering of the yeast Yarrowia lipolytica for the production of glycoproteins lacking the outer-chain mannose residues of N-glycans Appl. Environ. Microbiol. . 73 , PubMedID: 17513593, DOI:
- Aoki, K., Perlman, M., Lim, J.M., Cantu, R., Wells, L., Tiemeyer, M. 2007. Dynamic developmental elaboration of N-linked glycan complexity in the Drosophila melanogaster embryo J. Biol. Chem. . 282 , PubMedID: 17264077, DOI:
- Zhuo, Y., Chammas, R., Bellis, S.L. 2008. Sialylation of beta1 integrins blocks cell adhesion to galectin-3 and protects cells against galectin-3-induced apoptosis J. Biol. Chem. . 283 , PubMedID: 18676377, DOI:
- Fujita, N., Tamura, A., Higashidani, A., Tonozuka, T., Freeze, H.H., Nishikawa, A. 2008. The relative contribution of mannose salvage pathways to glycosylation in PMI-deficient mouse embryonic fibroblast cells FEBS J. . 275 , PubMedID: 18215164, DOI:
- Wagner-Rousset, E., Bednarczyk, A., Bussat, M.C., Colas, O., Corvaïa, N., Schaeffer, C., Van Dorsselaer, A., Beck, A. 2008. The way forward, enhanced characterization of therapeutic antibody glycosylation: comparison of three level mass spectrometry-based strategies J. Chromatogr. B Analyt. Technol. Biomed. Life Sci.. 872, PubMedID: 18672411, DOI:
- Graham, D.R., Mitsak, M.J., Elliott, S.T., Chen, D., Whelan, S.A., Hart, G.W., Van Eyk, J.E. 2008. Two-dimensional gel-based approaches for the assessment of N-Linked and O-GlcNAc glycosylation in human and simian immunodeficiency viruses Proteomics. 872, PubMedID: 19072736, DOI:
- Park, Y., Zhang, Z., Laremore, T.N., Li, B., Sim, J.S., Im, A.R., Ahn, M.Y., Kim, Y.S., Linhardt, R.J., Glycoconj, J. 2008. Variation of acharan sulfate and monosaccharide composition and analysis of neutral N-glycans in African giant snail (Achatina fulica) Glycoconjugate Journal. 25 , PubMedID: 18670878, DOI:
- Banala J. et al. 2008. Caged substrates for protein labeling and immobilization Chembiochem . 4 , PubMedID: 18033718, DOI:
- Baudino L, Shinohara Y, Nimmerjahn F, Furukawa J, Nakata M, Martínez-Soria E, Petry F, Ravetch JV, Nishimura S, Izui S. 2008. Crucial role of aspartic acid at position 265 in the CH2 domain for murine IgG2a and IgG2b Fc-associated effector functions J Immunol. . 181(9) , PubMedID: 18941257, DOI:
- Oh, D.B., Park, J.S., Kim, M.W., Cheon, S.A., Kim, E.J., Moon, H.Y., Kwon, O., Rhee, S.K., Kang, H.A. 2008. Glycoengineering of the methylotrophic yeast Hansenula polymorpha for the production of glycoproteins with trimannosyl core N-glycan by blocking core oligosaccharide assembly Biotechnol. J. . 3 , PubMedID: 18320567, DOI:
- Piro, J.R., Harris, B.T., Nishina, K., Soto, C., Morales, R., Rees, J.R., Supattapone, S. 2009. Prion protein glycosylation is not required for strain-specific neurotropism J Virol. . 83 , PubMedID: , DOI:
- Velho, A.M., Jarvis, S.M. 2009. Topological studies of hSVCT1, the human sodium-dependent vitamin C transporter and the influence of N-glycosylation on its intracellular targeting Exp. Cell Res. . 315 , PubMedID: 19379732, DOI:
- Boeggeman, E., Ramakrishnan, B., Pasek, M., Manzoni, M., Puri, A., Loomis, K.H., Waybright, T.J., Qasba, P.K. 2009. Site specific conjugation of fluoroprobes to the remodeled Fc N-glycans of monoclonal antibodies using mutant glycosyltransferases: application for cell surface antigen detection Bioconjug. Chem. . 20 , PubMedID: 19425533, DOI:
- Macnaughtan, M.A., Tian, F., Liu, S., Meng, L., Park, S., Azadi, P., Moremen, K.W., Prestegard, J.H. 2008. 13C-sialic acid labeling of glycans on glycoproteins using ST6Gal-I J. Am. Chem Soc. . 130 , PubMedID: 18700760, DOI:
- Gong, B., Cukan, M., Fisher, R., Li, H., Stadheim, T.A., Gerngross, T. 2009. Characterization of N-linked glycosylation on recombinant glycoproteins produced in Pichia pastoris using ESI-MS and MALDI-TOF Methods Mol. Biol. . 534 , PubMedID: , DOI:
- Rasmussen, T.N., Plenge, P., Bay, T., Egebjerg, J., Gether, U. 2009. A single nucleotide polymorphism in the human serotonin transporter introduces a new site for N-linked glycosylation Neuropharmacology . 57 , PubMedID: 19500602, DOI:
- Lee, K.J., Jung, J.H., Lee, J.M., So, Y., Kwon, O., Callewaert, N., Kang, H.A., Ko, K., Oh, D.B. 2009. High-throughput quantitative analysis of plant N-glycan using a DNA sequencer Biochem. Biophys. Res. Commun. . 380 , PubMedID: 19167352, DOI:
- Lee, K.J., Jung, J.H., Lee, J.M,, So, Y., Kwon, O., Callewaert, N., Kang, H.A., Ko, K., Oh, D.B. 2009. High-throughput quantitative analysis of plant N-glycan using a DNA sequencer Biochem. Biophys. Res. Commun. . 380 , PubMedID: 19167352, DOI:
- Gefter, J.V., Shaufl, A.L., Fink, M.P., Delude, R.L. 2009. Comparison of distinct protein isoforms of the receptor for advanced glycation end-products expressed in murine tissues and cell lines Cell Tissue Res. . 337 , PubMedID: 19415334, DOI:
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Learn about the core sequences and common modifications of N-linked and O-linked glycans in this video. Analysis of these glycans can be accomplished with the use of deglycosylation enzymes, which can provide complete sugar removal with no protein degradation.
Learn how glycosidases are used to analyze multiple glycoprotein samples. Here, the model glycoprotein, hCGβ, which carries both N- and O-glycans, is demonstrated.
Behind the Paper: An engineered Fbs1 carbohydrate binding protein for selective capture of N-glycans and N-glycopeptides
Minyong and Jim summarize their recent Nature Communications publication describing selective capture of N-glycans and N-glycopeptides by an engineered high affinity Fbs1 carbohydrate binding protein.
Learn more about the structural elucidation of complex and diverse heparin oligosaccharides using Bacteroides Heparinase I, II and III in combination with downstream mass spec analysis.
This one-step deglycosylation protocol is for use with New England Biolabs’ Rapid PNGase F. Learn more about Rapid PNGase F.
This two-step deglycosylation protocol is for use with New England Biolabs’ Rapid PNGase F. Learn more about Rapid PNGase F.