Specific Activity ＞140 U/mg;Activity ＞14 U/ml for sensitive chromatographic and analytical workflows requiring minimal baseline interference.
Store at 2-8°C Ships Wet ice Check lot-specific COA for exact specifications.
SDS, COA, datasheet, and spec sheet available for download. Lot-specific COA accessible via lot number lookup.
Cited in 0 peer-reviewed publications across chromatography, organic synthesis, and cross-coupling reactions.
Endo-beta-galactosidase cleaves internal ß(1-4) galactose linkages in unbranched, repeating poly-N-acetyllactosamine structures.
Product Description
Endo-Beta-Galactosidase cleaves internal β(1-4) galactose linkages in unbranched, repeating poly-N-acetyllactosamine structures. Sulfated structures such as keratan sulfate are also cleaved. Branching and/or fucosylation of the substrate may decrease or eliminate cleavage.
Endo-Beta-Galactosidase is useful for identifying and removing poly-N-acetyllactosamine structures on many biologically important glycoconjugates.
Source Recombinant from Bacteroides fragilis
Specificity Cleaves internal ß(1-4) galactose linkages in unbranched, repeating poly-N-acetyllactosamine [GlcNAc-ß(1-3)Gal-ß(1-4)]n structures are the preferred substrate. Sulfated structures such as keratan sulfate are also cleaved. Branching and/or fucosylation of the substrate may decrease or eliminate cleavage. Sulfation of C-6 on galactose will block cleavage. Oligosaccharidesof the neo-lacto group are cleaved at greatly educed rates depending on the deviation from the preferred substrate. For example, Gal-ß(1-3)GlcNAc-ß(1-3)Gal-ß(1-4)Glc is cleaved at 5×10-5 the rate of keratan sulfate. Specificity is similar to the Escherichia freundii enzyme. Specificity is similar to the Escherichia freundii enzyme except that it is limited to cleaving N-acetyllactosamine extensions on tetraantennary structures of erythropoietin.
Purity Endo-Beta-Galactosidase is tested for contaminating protease as follows: 10 μg of denatured BSA is incubated for 24 hours at 37°C with 2 μL of enzyme. SDS-PAGE analysis of the treated BSA shows no evidence of degradation.
EC 3.2.1.103
CAS 55072-01-0
Specific Activity >140 U/mg
Activity >14 U/ml
Molecular weight ~32,000 daltons
Optimum pH 5.8
Specific Activity
One unit of Endo-Beta-Galactosidase is defined as the amount that will liberate one µmole of reducing sugar per minute at 37˚C and pH 5.8 from bovine corneal keratan sulfate.
Contents
60 µl aliquot of enzyme (0.9 U) in 20 mM tris-HCl, pH 7.5
1 vial reaction buffer- 250mM Sodium phosphate, pH 5.8
Suggested usage
For glycoproteins:
1. Add up to 100 µg of glycoprotein to a tube.
2. Add 4 ul 5X buffer and water to 19 µl.
3. Add 1 µl enzyme.
4. Incubate at 37˚C for 2 hrs.
Procedure for oligosaccharides:
Same as above except incubate from several hours to several days depending on the substrate. Add bovine serum albumen to 2 mg/ml to stabilize the protein during extended incubations.
Storage
Stability Stable at least 24 months when stored properly. Several days exposure to ambient temperature will not reduce activity. Active for at least 5 days under reaction conditions.
The production strain of E. coli has been extensively tested and does not produce any detectable glycosidases.
Comprehensive hazard, handling, storage, and regulatory compliance document.
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