Recombinant SUMO Protease (Ulp1, His-tag), 2000 U

Product Information

Product Description/Introduction

Recombinant SUMO Protease is a highly active cysteinyl protease also known as Ulp1 (ubiquitin-like protein-specific protease 1).SUMO Protease cleaves in a highly specific manner, recognizing the tertiary structure of the ubiquitin-like (UBL) protein, SUMO rather than an amino acid sequence. The optimal temperature for cleavage is 30°C; however, the enzyme is active over wide ranges of temperature (2-30) and pH (pH 6.010.0). 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.SUMO Protease, a highly active cysteinyl protease also known as Ulp, is a recombinant fragment of Ulp1 (Ubl-specific protease 1) from Saccharomyces cerevisiae. SUMO Protease cleaves in a highly specific manner, recognizing the tertiary structure of the ubiquitin-like (UBL) protein, SUMO rather than an amino acid sequence. The protease can be used to cleave SUMO from recombinant fusion proteins. The optimal temperature for cleavage is 30°C; however, the enzyme is active over wide ranges of temperature (2-30) and pH (pH 6.010.0). Following digestion, SUMO Protease is easily removed from the cleavage reaction by affinity chromatography using the polyhistidine tag at the N-terminus of the protease.

Application: The Protease can be used to cleave SUMO from recombinant fusion proteins.

Source: Saccharomyces cerevisiae cysteine protease, It is purified from E. coli by affinity chromatography using the polyhistidine tag.

Enzyme activity: One unit of SUMO Protease cleaves 85% of 2µg control substrate in 1h at 30 and pH 8.0.

Inactivation or inhibition: Concentrations of the imidazole higher than 150 mM can adversely affect the activity of the 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 Shipping Conditions

Ship with wet ice; Store at -20; Valid for 12 months.

Product Components

Assay Protocol/Procedures

1. Protease digestion system recommended：

Note: For most fusion proteins, SUMO Protease functions optimally in a reaction mixture containing 150 mM NaCl; however,conditions may be optimized by varying the NaCl concentration from 0 mM to 300 mM.

2. Place at 30 for 1, 2 or 4 h. It can also digest at 4 overnight (about 16 h) if the target protein is not stable at 30.；

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

Note

1. Place in an ice box or ice bath when used, and separately store at -20 immediately after use.

2. For your safety and health, please wear safety glasses, gloves, or protective clothing.

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 are 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 addin the enzyme digestion system is 10:1)

For Research Use Only!

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