background:

Fucose is an important component of many oligosaccharide (O- or N-linked) and polysaccharide structures, as well as glycoproteins and glycolipids, which are often involved in various biological events, including fertilization, embryogenesis, apoptosis, signal transduction, and disease progression, such as rheumatoid arthritis, inflammation, cancer, and cystic fibrosis. Defucosylation of glycoconjugates is an important process to understand the biological effects of glycoconjugates .

α1-2- fucosidase is a highly specific exoglycosidase that catalyzes the hydrolysis of linear α1-2 linked L-fucosyl groups from oligosaccharides (linear substrates are defined as having no branches on adjacent residues).

Rhinogen ^® α1-2 Fucosidase is a recombinant glycosidase expressed in E. coli and purified by multi-step chromatography. Its molecular weight is about 97.7kD. It is active under neutral conditions and has the best reaction activity at 37°C. The reaction time can be appropriately extended depending on the digestion substrate.

Product Packaging:

Rhinogen ^® α1-2 Fucosidase (α 1-2 Fucosidase ) packaging specifications are as follows:

Supporting reagents:

The reagents provided with Rhinogen ® ^α1-2 Fucosidase are as follows:

Product Source:

Recombinantly expressed in E. coli, with a theoretical molecular weight of approximately 97.7KD and a 6×His tag at the C-terminus .

Product quality:

SDS-PAGE analysis showed that the purity was ≥95%; no contaminating exoglycosidase, endoglycosidase and protease activities were detected.

Product Features:

α1-2 fucosidase is a highly specific exoglycosidase that catalyzes the hydrolysis of α1-2 linked fucose on glycoproteins or free oligosaccharides.

Enzyme activity definition :

Rhinogen ^® α 1-2 Fucosidase hydrolyzes ≥90% of fucose from 11 nmol 2'-fucosyllactose in 20 mM Tris pH 6.8 at 37°C for 30 minutes .

Storage conditions:

1. Use ice packs for transportation. Please place the enzyme at 2-8℃ immediately after receiving the product; the supporting reagents should be stored at -20℃.

2. This product does not contain preservatives. Please ensure aseptic operation to avoid contamination.

application:

1. Glycan structure analysis;

2. Characterization and quality control of therapeutic recombinant proteins;

3. Eliminate glycoprotein heterogeneity.

characteristic:

Rhinogen ^® α1-2 Fucosidase is a highly purified and very stable exoglycosidase with the characteristics of high stability and compatibility with downstream assays.

1. High purity: no contaminating protease/other glycosidase, purity ≥95%;

2. High stability: Each batch of products undergoes strict quality control to achieve batch-to-batch stability ;

3. Compatible with downstream HPLC and mass spectrometry: It does not contain glycerol, and the enzyme digestion system is compatible with the HPLC and mass spectrometry workflow environment.

Recommended use:

1. Take 1µg of glycoprotein or 100nM oligosaccharide sample and add purified water to the reaction system to 9μl;

2. Add 1µl 10× Glyco Buffer 3;

3. Add 1-2 µl Rhinogen ^® α1-2 Fucosidase and mix gently;

4. Incubate at 37°C for 1 hour.

Note: Optimize enzyme concentration and incubation time appropriately depending on substrate type.

Instructions:

1. For different glycoprotein samples, it is necessary to experiment to find the most suitable enzyme concentration and reaction time;

2. The reaction can be scaled up or down linearly;

3. For different glycoprotein samples, the required enzyme amount needs to be properly optimized or the optimal operation method needs to be determined based on experience. In a 10~25μl reaction system, the recommended initial enzyme amount for 1μg of glycoprotein is 1~2μl, and the reaction is for 1 hour. If it is not completely digested, it is recommended to treat overnight.

Note:

This product is for research use only and is not intended for diagnostic or therapeutic use in humans or animals.