background:

O -glycoproteases are a growing group of highly specific endoproteases that recognize and cleave glycopeptides/glycoproteins adjacent to O -glycosylated serine or threonine. MS analysis of glycopeptides generated by O -glycoprotease cleavage provides important information about the O -glycosylation sites in the protein sequence and the structure of the O - glycans attached to each glycosylation site .

Overview:

Rhinogen ^® O- GlyCORTAR protease is recombinantly expressed in E. coli and contains a His tag with a theoretical molecular weight of approximately 97kDa. Rhinogen ^® O- GlyCORTAR protease is an O- glycan- dependent protease that digests proteins carrying mucin-type O- glycans, including sialylated substrates, and the N-termini of glycosylated Ser and Thr residues. The enzyme does not digest unmodified serine or threonine residues, nor does it digest N-glycosylation sites of glycoproteins; and it can digest a variety of O- glycan structures, including sialylated glycosylated core 1 and core 2 structures and Tn antigens.

Packing Specifications:

Rhinogen ^® O - GlyCORTARprotease packaging specifications are as follows:

Enzyme activity definition :

In the presence of 50 mM Tris-HCl, 150 mM NaCl, pH 7.6, at 37°C for 30 minutes, 1 unit of enzyme (1 U) can digest 30 µg of etanercept by ≥ 95%, as confirmed by SDS-PAGE.

Storage conditions:

The product is transported in ice packs. Please store it at 2~8℃ after receiving it. Do not freeze it.

Product Features:

1. High purity: SDS-PAGE analysis, purity ≥ 90%;

2. High specificity: only proteins carrying mucin-type O- glycans can be digested, including sialylated substrates, and N-termini of glycosylated Ser and Thr residues;

3. Wide applicability: accepts a variety of O -glycan structures, including sialylated glycosylated core 1 and core 2 structures and Tn antigens;

4. No animal origin: Recombinant production, no exogenous virus contamination, and no animal origin raw materials are used in the production process.

Product Application:

1. O -glycosylation analysis;

2. O -glycan site determination.

Antibody digestion:

1. Digestion was performed at a ratio of 1U Rhinogen ^® O - GlyCORTAR protease /1μg glycoprotein;

2. Incubate in 50 mM Tris-HCl, 150 mM NaCl, pH 7.6, 37°C for 2 hours.

Precautions for use:

1. For complex substrates, the incubation time can be extended to overnight and optimized according to the specific situation;

2. The appropriate final concentration of glycoprotein in the reaction system is 0.1-10 mg/ml;

3. This product is for research use only and is not intended for human or animal diagnosis or treatment.