3. Recombinant SUMO Protease (Ulp1, His-tag), 2000 U $655

Product Information

Description

Recombinant SUMO Protease (Small Ubiquitin-related Modifier Protease) is a common cysteine protease (Ubl-specific protease 1, UBL-specific Modifier). ubiquitin-like protein-specific protease 1, Ulp1). It can highly specifically recognize the tertiary structure of SUMO (Small Ubiquitin-Like Modifier) protein and hydrolyze the peptide bond after Gly-Gly in X-Gly-Gly-X (except Pro) peptide at the carboxyl end of SUMO protein. Therefore, the target protein released by the protease after cleavage of the fusion protein does not carry excess amino acids. The optimal pH and temperature of SUMO Protease digestion were 8.0 and 30℃, respectively. It has good stability, and has high enzyme activity in pH 6.0-10.0, temperature 2-30℃, and ionic strength 0-400 mM NaCl. In the actual operation, in order to preserve the structure and biological activity of the target protein, it is recommended to use Recombinant SUMO Protease at 4℃ overnight. At the same time, adding appropriate concentration of DTT (0.5-2 mM) in the digestion system can also significantly improve the digestion efficiency.

Application: It is often used to remove SUMO tag proteins from fusion proteins. The protease itself carries a His tag that binds to the nickel column.

Source: Saccharomyces cerevisiae cysteine protease, recombinant expressed in Escherichia coli with His tag.

Enzyme activity: It was defined as 1 active unit of enzyme required to cleave more than 85% of 2 μg substrate (containing SUMO-tagged protein) within 1 h at 30 ° C and pH8.0.

Inactivation or inhibition: higher than 150 mM imidazole affects the restriction efficiency of Recombinant SUMO Protease.

Purity and concentration: Purity ≥95% by SDS-PAGE, 10 U/μL.

Recombinant SUMO protease storage solution：25 mM Tris-HCl，pH 8.0，0.1% NP-40，250 mM NaCl，500 μM DTT，50% (v/v) glycerol.

10×SUMO Buffer+Salt：500 mM Tris-HCl，pH 8.0，2% NP-40，1.5 M NaCl，10 mM DTT.

10×SUMO Buffer – Salt：500 mM Tris-HCl，pH 8.0，2% NP-40，10 mM DTT.

Storage and Handling Conditions

Transport with wet ice; Store at -20℃, valid for 12 months.

Component

Assay Protocol

1. Protease digestion system recommended：

Note: For most fusion proteins, the optimal concentration of NaCl in SUMO Protease reaction buffer is 150 mM, and the concentration of NaCl (0-300 mM) can be adjusted according to the actual conditions.

2. The reaction system was placed at 30℃ for 1, 2 or 4 h. If the stability of the target protein is not good at 30℃, the reaction system can be considered to be digested at 4℃ overnight (about 16 h).；

3. 10 μL of enzyme digestion product was taken for SDS-PAGE electrophoresis to evaluate the amount of enzyme digestion and reaction conditions.

Note:

1. Enzyme products should be placed in an ice box or ice bath when used, and stored at -20℃ immediately after use. It is recommended to store them separately；

2. For your safety and health, please wear a lab coat and disposable gloves when operating.

Examples of enzyme digestion applications

Figure 1: Effect of recombinant SUMO Protease digestion of control substrate protein (SUMO+B fusion protein) (The enzyme digestion conditions were 30℃ and pH 8.0, respectively for 0 min, 10 min, 20 min, 30 min, 1 h, 2 h and 4 h, corresponding to Lane 1-7 in turn, and the mass ratio of substrate and enzyme added in the enzyme digestion system was 10:1)

For Research Use Only!