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Analysis of glycosphingolipids: recombinant endoglycoceramidase II (rEGCase II)
Recombinant endoglycoceramidase II (rEGCase II) is an endo-type enzyme that cleaves the linkages between oligosaccharides and ceramides of acidic and neutral glycosphingolipids, producing intact oligosaccharide and ceramide byproducts. This enzyme does not act on galactosyl- or glucosylceramides. It also does not hydrolyze cerebrosides, sulfatides, sphingomyelin, glycoglycerolipids, or glycoproteins. Globo-type glycosphingolipids are highly resistant to rEGCase II hydrolysis. rEGCase II is most suited to the structural analysis of glycosphingolipids.
- Structural analysis of glycosphingolipids
Recombinant E. coli
rEGCase is free from the following exoglycosidase activities: alpha-galactosidase, beta-galactosidase, alpha-N-acetylgalactosaminidase, beta-N-acetylgalactosaminidase, beta-N-acetyl-glucosaminidase, alpha-mannosidase, alpha-fucosidase, and sialidase. This enzyme is also free from glycopeptidase, endo-beta-N-acetyglucosaminidase, proteinase, and sphingomyelinase activities.
Store at –20°C until use. Thawed solution is stable for 2–3 days at 4°C.
This product contains detergents and requires them for optimal activity. Do not use on living cells.
- Systematic name: Oligoglycosylglucosylceramide glycohydrolase (EC 220.127.116.11)
- Form: 20 mM sodium acetate solution (pH 6.0), 0.2% BSA, 0.1% Lubrol PX
- Concentration: 100 mU/100 µl
- Optimum pH: 5.0–6.0
- Inhibitors: 1 mM Hg2+, Zn2+, Cu2+
- Not inhibited by Ba2+, Ca2+, Mg2+, or EDTA
Definition of activity
One unit of enzyme activity is defined as the amount needed to catalyze the hydrolysis of 1 µmol asialo GM1 per minute.
Ito, M. & Yamagata, T. Purification and characterization of glycosphingolipid-specific endoglycosidases (endoglycoceramidases) from a mutant strain of Rhodococcus sp. Evidence for three molecular species of endoglycoceramidase with different specificities. J. Biol. Chem. 264, 9510–9519 (1989).
Park, J.T. & Johnson, M.J. A submicrodetemination of glucose. J. Biol. Chem. 181, 149–151 (1949).
Additional product information
Please see the product's Certificate of Analysis for information about storage conditions, product components, and technical specifications. Please see the Kit Components List to determine kit components. Certificates of Analysis and Kit Components Lists are located under the Documents tab.
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