background:

β1-4 Galactosidase is an important enzyme for glycosylation research. Rhinogen ^® β1-4 Galactosidase is a recombinant enzyme that is recombinantly expressed in E. coli BL21 and is capable of catalyzing the hydrolysis of β1-4 linked galactose residues at the non-reducing end of oligosaccharides or glycoproteins, as shown in Figure 1.

Figure 1 The cleavage site of β1-4 Galactosidase

Packing Specifications:

Rhinogen ^® β1-4 Galactosidase packaging specifications are as follows:

Storage system:

QPF-006 is supplied in liquid form in a buffer consisting of 50 mM NaCl, 20 mM Tris-HCl, 1 mM EDTA (pH 7.5, 25°C).

Supporting reagents:

The supporting reagents provided by Rhinogen ^{® β1-4 Galactosidase are as follows:}

Product Source:

Rhinogen ^® β1-4 Galactosidase is a recombinant enzyme that is produced by recombining the β1-4 Galactosidase gene and expressing it in E. coli BL21. Its molecular weight is approximately 94KD.

Product quality:

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

Product Features:

The optimal reaction pH is 6.0. Inactivation conditions: 65°C for 10 min.

Enzyme activity definition :

One unit of enzyme activity is defined as the amount of enzyme required to catalyze the hydrolysis of 1 μmol of o-nitrophenyl-β-D - galactopyranoside in 1 minute at 37°C and pH 5.5.

1U Rhinogen ^® β1-4 Galactosidase =1 U Prozyme Glyko ^® β(1-4)-Galactosidase. For the conversion of enzyme activity units of products from different companies, please refer to the activity unit conversion table as follows:

characteristic:

Rhinogen ^® β1-4 Galactosidase is a highly purified and very stable exoglycosidase with high stability and specific activity, suitable for proteomics and glycobiology research.

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

2. High stability: Each batch of β1-4 Galactosidase products undergoes strict quality control to achieve high stability;

3. High specific activity: efficient and complete release of all non-reducing terminal β1-4 linked galactose residues.

application:

1. Glycan structure analysis;

2. Characterization and quality control of therapeutic recombinant proteins;

3. Eliminate glycoprotein heterogeneity.

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 1;

3. Add 1µl Rhinogen ^® β1-4 Galactosidase and mix gently;

4. Incubate at 37°C for 1 hour.

Instructions:

1. The reaction system can be scaled up or down linearly;

2. For different glycoprotein samples, appropriate optimization or the best operation method needs to be determined based on experience. In a 10-25μl reaction system, for 1μg of glycoprotein, the recommended initial enzyme amount is 1-2μl and the reaction is for 1 hour. If it is not completely digested, it is recommended to treat overnight;

3. pNP -βGal is an effective positive control substrate.

4. If the molecular size of native and de-galactose residues of glycoproteins is different enough on the gel to be distinguished, SDS-PAGE can be used to evaluate the degree of glycosylation and deglycosylation of glycoproteins ;

5. If you want to separate and recover the glycans cleaved by β1-4 Galactosidase, the substrate concentration of glycoprotein should be about 10-20μM, and the reaction time should be appropriately extended;

6. This product is for research use only and is not intended for diagnosis or treatment of humans or animals.