Endo D
Product informationCode | Name | Size | Quantity | Price | |
---|---|---|---|---|---|
P0742S |
Endo D |
1500 units ( 50000 units/ml ) | - | Unavailable in your region | |
P0742L |
Endo D |
7500 units ( 50000 units/ml ) | - | Unavailable in your region |
Endo D
Product Introduction
Endo D is an endoglycosidase that cleaves within the chitobiose core of paucimannose N-linked glycans, with or without extensions in the antennae, from glycoproteins and glycopeptides.
- Tagged with a chitin binding domain (CBD) for easy removal
- Recombinant enzyme with no detectable exooglycosidase or other endoglycosidase contaminating activities
- Glycerol -free for optimal performance in HPLC and mass spectrometry analysis
- ≥95% purity, as determined by SDS-PAGE and intact ESI-MS
- Optimal activity and stability for up to 12 months
Catalog # | Size | Concentration |
---|---|---|
P0742S | 1500.0 units | 50000 units/ml |
P0742L | 7500.0 units | 50000 units/ml |
- Product Information
- Protocols, Manuals & Usage
- Tools & Resources
- FAQs & Troubleshooting
- Citations & Technical Literature
- Quality, Safety & Legal
- Other Products You May Be Interested In
Product Information
Description
Endo D, also known as
Endoglycosidase D, is a recombinant glycosidase,
which cleaves within the chitobiose core of
paucimannose N-linked glycans, with or without
extensions in the antennae. Endo D
is tagged with a chitin binding domain (CBD) for
easy removal from a reaction, and is supplied
glycerol-free for optimal performance in HPLC and
MS- intensive methods.
Product Source
A truncated Endo D gene cloned from Streptococcus pneumoniae and expressed in E. coli as a fusion to chitin binding domainSpecificity
Detailed Specificity- This product is related to the following categories:
- Endoglycosidases Products,
- Proteome Analysis Products,
- This product can be used in the following applications:
- Expression Systems,
- Glycan Sequencing,
- Proteomics,
- Recombinant Glycoprotein Expression, Glycoprotein Analysis
Reagents Supplied
Reagents Supplied
The following reagents are supplied with this product:
NEB # | Component Name | Component # | Stored at (°C) | Amount | Concentration | |
---|---|---|---|---|---|---|
Properties & Usage
Unit Definition
One unit is defined as the amount of enzyme required to remove > 95% of the carbohydrate from 10 μg of glycosidase-trimmed (trimannosyl core) Fetuin in 1 hour at 37°C in a total reaction volume of 10 μl.Reaction Conditions
1X GlycoBuffer 2
Incubate at 37°C
1X GlycoBuffer 2
50 mM Sodium Phosphate
(pH 7.5 @ 25°C)
Storage Buffer
20 mM Tris-HCl
50 mM NaCl
1 mM EDTA
pH 7.5 @ 25°C
Heat Inactivation
65°C for 10 minutesMolecular Weight
Apparent: 140000 daltonsUnit Assay Conditions
10 μg of glycosidase-trimmed (trimannosyl core) Fetuin are denatured with 1X DTT at 95°C for 3 minutes. After the addition of 1X GlycoBuffer 2, two-fold dilutions of Endo D are added and the reaction mix is incubated for 1 hour at 37°C. Separation of reaction products is visualized by SDS-PAGE.Related Products
Companion Products
Product Notes
- To deglycosylate a native glycoprotein, longerincubation time, as well as more enzyme, maybe required.
- Endo D is not recommended foruse with Glycoprotein Denaturing Buffercontaining both SDS and DTT, as Endo D is inhibited by SDS and, unlike otherendoglycosidases, NP-40 does not counteractthe SDS inhibition.
- Removal of Endo D from thedeglycosylation reaction can be scaled uplinearly with larger amounts of chitin magneticbeads.
- Chitin Magnetic Beads Binding Capacity is0.4 μg/μl of CBD-tagged protein.
- 50 μl of chitin slurry are sufficient to remove 1-5 μl of Endo D (50-250 units)
References
- McLeod, E., Shi, S. and Magnelli, P., New England Biolabs, Inc., unpublished results. Unpublished observation
Protocols, Manuals & Usage
Protocols
Tools & Resources
Selection Charts
FAQs & Troubleshooting
FAQs
- What is the tag on Endo D?
- What is the difference between PNGase F, Endo S and Endo D?
- What is a typical reaction protocol for Endo D?
- Will SDS inhibit Endo D?
- How do I eliminate Endo D from a reaction?
- What is the binding capacity of the Magnetic Chitin Beads used to remove Endo D?
- Is Endo D compatible with downstream analysis such as HPLC and Mass Spectrometry?
- 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?
- Can Endo D cleave glycoproteins under native conditions?
Citations & Technical Literature
Citations
Additional Citations
Quality, Safety & Legal
Quality Assurance Statement
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]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]- P0742S_L_v1_0011606
- P0742S_L_v1_0011603
- P0742S_L_v1_0011612
- P0742S_L_v1_0011706
- P0742S_L_v1_0011804
- P0742L_v2_10039713
- P0742S_v2_10039419
- P0742L_v2_10053304
- P0742S_v2_10053306
- P0742S_v2_10068257
- P0742L_v2_10065141
- P0742L_v2_10080275
- P0742L_v2_10084156
- P0742L_v2_10093423
- P0742S_v2_10084153
- P0742S_v2_10107166
- P0742L_v2_10096503
- P0742S_v2_10116016
- P0742L_v2_10116017
- P0742L_v2_10143868
- P0742S_v2_10143865
- P0742L_v2_10151217
- P0742L_v2_10167450
- P0742S_v2_10167451
- P0742S_v2_10191225
- P0742S_v2_10223135
- P0742L_v2_10191224
- P0742S_v2_10234752
- P0742L_v2_10235980
- P0742L_v2_10252797
- P0742S_v2_10254370
Safety DataSheets
The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.Endo D
DTT
GlycoBuffer 2
Legal and Disclaimers
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.
Other Products You May Be Interested In
The supporting documents available for this product can be downloaded below.