Catalog # E0577 was discontinued on November 23, 2020
The N-Glycan Sequencing Kit consists of seven well characterized, highly pure, recombinant exoglycosidase enzymes selected to simplify the process of characterizing typical N-linked glycan structures.
Recombinant enzymes with no detectable endoglycosidase or other exoglycosidase contaminating activities
Compatible with N-linked glycans released from a variety of CHO and murine derived antibodies, as well as N-linked glycans released from other glycoproteins
Simultaneous digestion with other exoglycosidases
Optimal activity and stability for up to 12 months
Storage Temperature: 4°C
Product Information
The N-Glycan Sequencing Kit consists of seven well characterized, highly pure, recombinant exoglycosidase enzymes selected to simplify the process of characterizing typical N-linked glycan structures. The extreme diversity of glycan structures on a glycoprotein makes elucidation of the profile challenging and often a number of orthogonal approaches are employed to verify the individual structures. The use of sequential or tandem exoglycosidase digestion of oligosaccharides followed by mass spectrometry or capillary electrophoresis analysis provides detailed carbohydrate sequence information and resolves ambiguities. The exoglycosidases are all active in a universal buffer system, GlycoBuffer 1, and can be used in single digests or in combination.
This kit is compatible with N-linked glycans released from a variety of CHO and murine derived antibodies, as well as N-linked glycans released from other glycoproteins. Optimal incubation times and enzyme concentration may vary for more complex glycans. The recommended enzyme quantity is sufficient for digestion of up to 130 pmol of released glycans (equivalent to released glycans from approximately 10 µgs of antibody) labeled with 2AA, 2AB or procainamide. Other commercially available “Instant labels” may require a larger quantity of enzyme or a longer incubation time for complete digestion.
Figure 1: Glycan Sample Preparation Workflow
Figure 2: N-Glycans released from Infliximab, labelled with procainamide, digested for 3 hours with exoglycosidases
Digestion of Infliximab with a sequential panel of exoglycosidases serves as a tool to elucidate and verify glycan profiles. Refer to the product manual for reaction conditions.
This product is related to the following categories:
Remove all traces of Acetonitrile before digesting labeled substrate with exoglycosidases for optimal results.
Use PCR tubes to avoid evaporation when incubating reactions overnight.
Include ZnCl2 in reactions with α1-2,3,6 Mannosidase (P0768) for optimal digestion.
Fucosidase O (P0749) requires an 18 hr incubation for complete digest of some instant labelled glycans and complex oligosaccharides with multiple branches.
If using Fucosidase O (P0749) alone, incubation temperature can be increased to 50°C for more efficient digestion.
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 & Change Notifications
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]
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]
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.
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