The large size of this product was discontinued on June 15, 2025. The small size will remain available.
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 24 months
Product Information
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 domain
Specificity
Detailed SpecificityFigure 1: Detailed specificity of Endo D The reaction contained 0.08 mU of α1-2 Mannosidase (Prozyme #GKX-5009), 50 units of Endo D and 2 μl 0.5 M Na0Ac pH 5.0 buffer in a total reaction volume of 10 μl. Reactions were incubated at 37°C for 2 hours. Following removal of the bottom branch, Endo D is active on a complex upper arm (1).
This product is related to the following categories:
The following reagents are supplied with this product:
NEB #
Component Name
Component #
Stored at (°C)
Amount
Concentration
P0742S
-20
Endo D
P0742SVIAL
-20
1 x 0.03 ml
50,000 units/ml
DTT
B0706SVIAL
-20
1 x 1 ml
0.4 M
GlycoBuffer 2
B3704SVIAL
-20
1 x 1 ml
10 X
P0742L
-20
Endo D
P0742LVIAL
-20
1 x 0.15 ml
50,000 units/ml
DTT
B0706SVIAL
-20
1 x 1 ml
0.4 M
GlycoBuffer 2
B3704SVIAL
-20
1 x 1 ml
10 X
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.
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.
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)
Recommended storage temperature is -20 °C.
For short-term use (within a few days), store at 4°C.
For long-term use (more than a few days), prepare aliquots and store at -20 °C to minimize freeze-thaw cycles.
References
McLeod, E., Shi, S. and Magnelli, P., New England Biolabs, Inc., unpublished results. Unpublished observation
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
Effective July 7, 2025, shelf life and storage recommendation updated.
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