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α1-2,4,6 Fucosidase O 收藏
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识别位点
- isoschizomers |
- compatible ends |
- single letter code
特性
Recombinant enzyme with no detectable endoglycosidase or other exoglycosidase contaminating
activities
Glycerol -free for optimal performance in HPLC and mass spectrometry analysis
≥95% purity, as determined by SDS-PAGE and intact ESI-MS
概述
α1-2,4,6 Fucosidase O is a broad specificity exoglycosidase that catalyzes the hydrolysis of terminal
α1-2, α1-4 and α1-6 linked fucose residues from oligosaccharides. α1-2,4,6 Fucosidase O cleaves
α1-6 fucose residues more efficiently than other linkages.
来源
Cloned from Omnitrophica bacterium and expressed in E. coli (1).
反应条件
1X GlycoBuffer 1. Incubate at 37°C.
Heat Inactivation: 65°C for 10 min
随酶提供的试剂
10X GlycoBuffer 1
1X GlycoBuffer 1
5 mM CaCl2
50 mM sodium acetate
(pH 5.5 @ 25°C)
分子量
Apparent: 49 kDa
单位定义
One unit is defined as the amount of enzyme required to cleave > 95% of the fucose from 1 nmol of G0F
from human IgG [GlcNAcβ1-2Manα1-6(GlcNAcβ1-2Manα1-3)Manβ1-4GlcNAcβ1-4GlcNAc(Fucα1-6)-
AMAC], in 1 hour at 37°C in a total reaction volume of 10 µl.
贮存温度
4°C
注意事项
1. Reactions may be scaled-up linearly to accommodate larger reaction volumes.
2. The enzymatic unit definition is calculated using a 37°C reaction temperature, to accommodate for
simultaneous glycosidase digestions and N-glycan sequencing protocols. However, the optimal
reaction temperature for α1-2,4,6 Fucosidase O is 50°C. A 37°C reaction temperature results in
~35% lower activity.
3. In order to achieve complete removal of core α1-6 fucose residues from glycans with instant labels,
it may be necessary to use longer incubation times (18 hours).
4. α1-2,4,6 Fucosidase O is only active on an intact antibody if it is trimmed to the trimannosyl core.
参考文献
1. Vainauskas, S., New England Biolabs, Inc. Unpublished observation
2. Wong-Madden, S.T. and Landry, D. (1995). Glycobiology. 5, 19-28.