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Exoglycosidases >
α1-2,3 Mannosidase |
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Prices are in US dollars and valid only for US orders.
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Substrate Specificity:
Description:
α1-2,3 Mannosidase is a highly specific exoglycosidase that catalyzes the hydrolysis of α1-2 and α1-3 linked D-mannopyranosyl residues from oligosaccharides (1).
Detailed Specificity: Specificity can vary depending on incubation time and concentration of substrate (Fig. 1).
Figure 1: Detailed specificity of α-1,2-3 Mannosidase. All reactions contained 32 units of α-1,2-3 Mannosidase and 1X G6 reaction buffer in a total reaction volume of 10 µl. Reactions were incubated at 37°C.
Source:
Cloned from Xanthomonas manihotis and expressed in E. coli (2).
Reagents Supplied:
G6 Reaction Buffer (10X)
BSA (100X)
Enzyme Properties
Heat Inactivation:
No
Molecular Weight:
Apparent: 90 kDa
Reaction & Storage Conditions
Reaction Conditions:
1X G6 Reaction Buffer
Supplemented with 100 μg/ml Bovine Serum Albumin
Incubate at
37°C.
1X G6 Reaction Buffer:
50 mM sodium acetate
5 mM CaCl2
pH 5.5 @ 25°C
Unit Definition:
One unit is defined as the amount of enzyme required to cleave > 95% of the non-reducing terminal α-D-mannose from 1 nmol Manα1-3Manβ1-4GlcNAc-7-amino-4-methyl-coumarin (AMC), in 1 hour at 37°C in a total reaction volume of 10 µl.
Unit Definition Assay: Two fold serial dilutions of α1-2,3 Mannosidase are incubated with 1 nmol AMC-labeled substrate in 1X G6 Buffer, supplemented with 100 µg/ml BSA, in a 10 µl reaction. The reaction mix is incubated for 1 hour at 37°C. Separation of reaction products are visualized via thin layer chromatography (1).
Concentration:
32,000 units/ml
Storage Conditions:
20 mM Tris-HCl
50 mM NaCl
1 mM Na2EDTA
pH 7.5 @ 25°C
Storage Temperature:
4°C
Notes
General notes:- p-nitrophenyl-α-D-mannopyranoside is NOT a substrate for this enzyme
FAQs
- How much exoglycosidase should be used?
- What is a good positive control for this α1-2, 3 Mannosidase?
- Do detergents inhibit exoglycosidases/endoglycosidases?
- Do detergents inhibit exoglycosidases/endoglycosidases?
- What are Glycosidases and their uses?
Quality Control for Current Lot
Quality control values for a specific lot can be found on the datacard which accompanies each product.
Quality Assurance Statement:
No contaminating exoglycosidase or proteolytic activity could be detected.
Glycosidase Assays:
32 units of α1-2,3 Mannosidase were incubated with 0.1 mM of flourescently-labeled oligosaccharides and glycopeptides, in a 10 µl reaction for 20 hours at 37°C. The reaction products were analyzed by TLC for digestion of substrate.
No other glycosidase activities were detected (ND) with the following substrates:
β-N-Acetylglucosaminidase:
GlcNAcβ1-4GlcNAcβ1-4GlcNAc-AMC ND
α-N-Acetylgalactosaminidase:
GalNAcα1-3(Fucα1-2)Galβ1-4Glc-AMC ND
α-Fucosidase:
Fucα1-2Galβ1-4Glc-AMC ND
Galβ1-4 (Fucα1-3)GlcNAcβ1-3Galβ1-4Glc-AMC ND
β-Galactosidase:
Galβ1-3GlcNAcβ1-4Galβ1-4Glc-AMC ND
Galβ1-4GlcNAcβ1-2Manα1-6Manβ1-4GlcNAc-AMC ND
α-Galactosidase:
Galα1-3Galβ1-4Galα1-3Gal-AMC ND
α-Mannosidase:
Manα1-6Manα1-6(Manα1-3)Man-AMC ND
α-Neuraminidase:
Neu5Acα2-3Galβ1-3GlcNAcβ1-3Galβ1-4Glc-AMC ND
β-Glucosidase:
Glcβ1-4Glcβ1-4Glc-AMC ND
α-Glucosidase:
Glcα1-6Glcα1-4Glc-AMC ND
β-Xylosidase:
Xylβ1-4Xylβ1-4Xylβ1-4Xyl-AMC ND
β-Mannosidase:
Manβ1-4Manβ1-4Man-AMC ND
Endo F1, F2, H:
Dansylated invertase high mannose. ND
Endo F2, F3:
Dansylated fibrinogen biantennary. ND
PNGase F:
Fluoresceinated fetuin triantennary. ND
Protease Assay::
After incubation of 220 units of α1-2,3 Mannosidase with 0.2 nmol of a standard mixture of proteins for 20 hours at 37°C, no proteolytic activity could be detected
by SDS-PAGE.
References
- Wong-Madden, S.T. and Landry, D. (1995) Glycobiology, 5, 19-28.
- Guthrie, E.P. and Taron, C.H., New England Biolabs, unpublished observations.
Reagents Sold Separately
BSA
Companion Products
Exoglycosidase Reaction Buffer Pack
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