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β-N-Acetyl-hexosaminidasef > FAQ |  | β-N-Acetyl-hexosaminidasef FAQ
See the Protein Tools FAQ also.
Q1: How much exoglycosidase should be used?
Q2: Do detergents inhibit exoglycosidases/endoglycosidases?
Q3: What is a good positive control for β-N-Acetylhexosaminidasef?
Q4: Do detergents inhibit exoglycosidases/endoglycosidases?
Q5: What are Glycosidases and their uses?
Q1: How much exoglycosidase should be used?
A1: The amount of enzyme required varies when different substrates are used. Start with 1-2 µl for 1µg of glycoprotein or 100 nM of oligosacharide for one hour in a 10-25 µl reaction. If there is still undigested material, let the reaction go overnight.
Q2: Do detergents inhibit exoglycosidases/endoglycosidases?
A2: At moderate levels (0.5-1.0% ionic and non-ionic detergents) most of the glycosidases show satisfactory activity or are unaffected. One major exception is PNGase F. SDS inhibits it.
Q3: What is a good positive control for β-N-Acetylhexosaminidasef?
A3: pNP-GlcNAc
Q4: Do detergents inhibit exoglycosidases/endoglycosidases?
A4: At moderate levels (0.5-1.0% ionic and non-ionic detergents) most of the glycosidases show satisfactory activity or are unaffected. One major exception is PNGase F as it is inhibited by SDS.
Q5: What are Glycosidases and their uses?
A5: Glycosidases are used to get information about the carbohydrate groups attached to glycoproteins and glycopeptides. They come in two varieties, endoglycosidases that cleave entire carbohydrate groups from proteins and exoglycosidases that remove monosaccharides from the nonreduced ends of the carbohydrate. A reduced end is one generated by an endoglycosidase. Researchers frequently use an endoglycosidase followed by one or more exoglycosidases and then analyze the products using SDS-PAGE or various types of liquid chromatography.
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