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324716 Endo-α-N-acetylgalactosaminidase, Streptococcus pneumoniae, Recombinant, E. coli

Endo-α-N-acetylgalactosaminidase, Streptococcus pneumoniae, Recombinant, E. coli, CAS 59793-96-3, catalyzes the hydrolysis of the unsubstituted Galβ1,3GalNAc core disaccharide attached to Ser or Thr.

MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.

Synonyms: O-Glycopeptide endo-D-galactosyl-N-acetyl-α-galactosaminohydrolase, O-Glycosidase

324716
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Overview

Replacement Information

Products

Catalogue NumberPackaging Qty/Pack
324716-50MIU Plastic ampoule 50 miu
Description
OverviewRecombinant, Streptococcus pneumoniae Endo-α-N-acetylgalactosaminidase expressed in E. coli. Catalyzes the hydrolysis of the unsubstituted Galβ1,3GalNAc core disaccharide attached to serine or threonine residues of glycopeptides and glycoproteins to afford free oligosaccharides. For carbohydrates containing sialic acid or fucose, pretreatment with neuraminidase or fucosidase is required.
Note: 1 mU = 1 milliunit.
Catalogue Number324716
Brand Family Calbiochem®
SynonymsO-Glycopeptide endo-D-galactosyl-N-acetyl-α-galactosaminohydrolase, O-Glycosidase
References
ReferencesWang, A.M., et al. 1998. Mol. Genet. Metab. 65, 165.
Iwase, H., and Hotta, K. 1993. Methods Mol. Biol. 14, 151.
Fan, J.Q., et al. 1990. Agric. Biol. Chem. 54, 233.
Umemoto, J., et al. 1978. Anal. Biochem. 91, 186.
Glasgow, L.R., et al. 1977. J. Biol. Chem. 252, 8615.
Product Information
CAS number59793-96-3
Activity≥1 unit/ml
Unit of DefinitionOne unit is defined as the amount of enzyme that will catalyze the release of 1.0 µmol p-nitrophenol from p-nitrophenyl-2-acetamido-2-deoxy-3-O-(β-D-galactopyranosyl)-α-D-galactopyranoside per min at 37°C, pH 5.0.
EC number3.2.1.97
FormLiquid
FormulationIn 50 mM sodium phosphate buffer, pH 7.5.
Quality LevelMQ100
Applications
Biological Information
Specific Activity≥10 units/mg protein
Physicochemical Information
ContaminantsN-acetylglucosaminidase, α- and β-galactosidase, α-mannosidase, neuraminidase, proteases: none detected
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
Product Usage Statements
Storage and Shipping Information
Ship Code Blue Ice Only
Toxicity Standard Handling
Storage +2°C to +8°C
Do not freeze Yes
Packaging Information
Transport Information
Supplemental Information
Specifications
Global Trade Item Number
Catalogue Number GTIN
324716-50MIU 04055977215977

Documentation

Endo-α-N-acetylgalactosaminidase, Streptococcus pneumoniae, Recombinant, E. coli SDS

Title

Safety Data Sheet (SDS) 

Endo-α-N-acetylgalactosaminidase, Streptococcus pneumoniae, Recombinant, E. coli Certificates of Analysis

TitleLot Number
324716

References

Reference overview
Wang, A.M., et al. 1998. Mol. Genet. Metab. 65, 165.
Iwase, H., and Hotta, K. 1993. Methods Mol. Biol. 14, 151.
Fan, J.Q., et al. 1990. Agric. Biol. Chem. 54, 233.
Umemoto, J., et al. 1978. Anal. Biochem. 91, 186.
Glasgow, L.R., et al. 1977. J. Biol. Chem. 252, 8615.
Data Sheet

Note that this data sheet is not lot-specific and is representative of the current specifications for this product. Please consult the vial label and the certificate of analysis for information on specific lots. Also note that shipping conditions may differ from storage conditions.

Revision19-June-2008 RFH
SynonymsO-Glycopeptide endo-D-galactosyl-N-acetyl-α-galactosaminohydrolase, O-Glycosidase
DescriptionRecombinant, Streptococcus pneumoniae Endo-α-N-acetylgalactosaminidase expressed in E. coli. Catalyzes the hydrolysis of the unsubstituted Galβ1,3GalNAc core disaccharide attached to serine or threonine residues of glycopeptides and glycoproteins to afford free oligosaccharides.
FormLiquid
FormulationIn 50 mM sodium phosphate buffer, pH 7.5.
Recommended reaction conditions
Endo-α-N-acetylgalactosaminidase, Streptococcus pneumoniae, Recombinant, E. coli Note: This protocol is provided only as a general guidline. Researchers should standardize this assay for their own specific needs and should consult published literature. Endo-α-N-acetylgalactosaminidase cleaves only unsubstituted Galβ1,3GalNAc disaccharides attached to the serine or threonine residues of glycoproteins or glycopeptides. Substitutions such as sialic acid, galactose, fucose, or N-acetylglucosamine must first be removed with the appropriate exoglycosidase prior to treatment with Endo-α-N-acetylgalactosaminidase. At a minimum, a neuraminidase (e.g. α2-3,6,8-Neuraminidase, Arthrobacter ureafaciens, Recombinant, E. coli, Cat. No. 480716) is almost always required to remove sialic acid. Required Reagents • 250 µM substrate (p-nitrophenol-2-acetamido-2-deoxy-3-O-(β-Dgalactopyranosyl)-α-D-galactopyranoside (Cat. No. 575303; also called T antigen) in 5 mM Sodium phosphate, pH 5.0 • 1 M Na2CO3 • Spectrophotometer Protocol 1. Adjust the spectrophotometer to read 400 nm. 2. Add 0.4 ml of substrate solution to two tubes and warm to 37°C. 3. Add 5 µl of enzyme to one tube and mix. Continue incubating at 37°C. 4. Following a 5 minute incubation, add 0.6 ml of 1 M Na2CO3 to both tubes. 5. Blank the spectrophotometer using the control tube without enzyme. 6. Read the absorbance of the sample at 400 nm. 7. Calculate the units: Units/ml = A/18.3 TV A = absorbance of the sample T = time in min V = volume of enzyme in ml 18.3 = millimolar extinction coefficient of pNP at pH >10 Note: Endo-α-N-acetylgalactosaminidase cleaves methylumbelliferyl-α-D-Nacetylgalactosamide (but not pNP-α-D-N-acetylgalactosamide) at 0.1% the rate of p-nitrophenyl-2-acetamido-2-deoxy-3-O-(β-D-galactopyranosyl)-α-D-galactopyranoside. Assay for Deglycosylation Required Reagents • Endo-α-N-acetylgalactosaminidase • 5X assay buffer: 250 mM sodium phosphate, pH 5.0 • sterile dH2O • α2-3,6,8-Neuraminidase, Arthrobacter ureafaciens, Recombinant, E. coli, (Cat. No. 480716) • glycoprotein sample • SDS-PAGE system Protocol 1. Add up to 100 µg of glycoprotein to a tube. 2. Add water to 13 µl and then add 4 µl of 5X assay buffer. 3. Add 1 µl of α2-3,6,8-Neuraminidase, Arthrobacter ureafaciens, Recombinant, E. coli. 4. Add 2 µl of Endo-α-N-acetylgalactosaminidase 5. Incubate at 37°C for 1 h. Cleavage may be monitored by SDS-PAGE if the size differential between the native and the de-O-glycosylated protein is sufficient for detection.
CAS number59793-96-3
EC number3.2.1.97
ContaminantsN-acetylglucosaminidase, α- and β-galactosidase, α-mannosidase, neuraminidase, proteases: none detected
Specific activity≥10 units/mg protein
Activity≥1 unit/ml
Unit definitionOne unit is defined as the amount of enzyme that will catalyze the release of 1.0 µmol p-nitrophenol from p-nitrophenyl-2-acetamido-2-deoxy-3-O-(β-D-galactopyranosyl)-α-D-galactopyranoside per min at 37°C, pH 5.0.
Storage +2°C to +8°C
Do Not Freeze Yes
Toxicity Standard Handling
ReferencesWang, A.M., et al. 1998. Mol. Genet. Metab. 65, 165.
Iwase, H., and Hotta, K. 1993. Methods Mol. Biol. 14, 151.
Fan, J.Q., et al. 1990. Agric. Biol. Chem. 54, 233.
Umemoto, J., et al. 1978. Anal. Biochem. 91, 186.
Glasgow, L.R., et al. 1977. J. Biol. Chem. 252, 8615.