Recombinant SUMO Protease (Ulp1, His-tag)

 

Product Name	Cat. No.	Spec.
Recombinant SUMO Protease (Ulp1, His-tag)	G3411-2000U	2000 U

 

 

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 6.0–10.0). 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 6.0–10.0). 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.

 

 

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.

 

 

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

 

 

Component Number	Component	G3411-2000U
G3411-1	Recombinant SUMO Protease (His-tag) (10 U/μL)	200 μL
G3411-2	10×SUMO Buffer + Salt	1 mL
G3411-3	10×SUMO Buffer - Salt	1 mL
G3411-4	Control Protein	100 μL
Manual

 

 

1. Protease digestion system recommended：

Component	Volume
10×SUMO Buffer +/- Salt	5 μL
Target Proteins	20 μg
Recombinant SUMO Protease (His-tag) (10 U/μL)	1 μL
ddH2O	To 50 μL

 

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.

 

 

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!