Description
SUMO protease recognizes the complete SUMO (Small Ubiquitin-like Modifier) label protein with 100 amino acids and efficiently cuts the SUMO from the fusion protein. Compared with the recognition sites of EK and TEV, SUMO protease has high specificity due to its long recognition sequence, and maintains high activity in a wide range of reaction environmental systems, such as temperature (4-30℃), pH (5.5-9.5), etc. SUMO protease also has a polymerized His label, which is convenient to remove SUMO protease and purify the target protein by affinity chromatography.
Specifications
|
Cat.No. |
20410ES60 / 20410ES70 / 20410ES80 |
|
Size |
100 U / 500 U / 1000 U |
|
Enzyme activity |
1 U/μL |
|
Unit Definition |
The amount of enzyme required to cut >85% of the 3 μg substrate in a 1×rTEV Buffer (50 mM Tris, pH 8.0, 0.1 mM EDTA, 1 mM DTT) at 30℃ for 1 h was defined as a unit of activity |
|
Source |
Expressed in E. coli |
|
Label |
Polymerized His label |
|
Protein purity |
> 90% |
|
Molecular weight |
26.55 kDa |
|
Storage Buffer |
25 mM Tris-HCl, pH 8.0、0.1% Igepal (NP-40)、250 mM NaCl、500 μM DTT、50% glycerin |
Components
Note：The enzyme activity definition uses 10×Protease Buffer (-salt). SUMO Protease has relatively high activity in 10×Protease Buffer (-salt), but for unstable substrates, 10×Protease Buffer (+salt) should be selected. We also provide reaction buffer 20410-B2 with +salt. Choose the correct buffer for your experiment. 20410-B2：500 mM Tris-HCl, pH 8.0, 2% Igepal (NP-40), 1.5 M NaCl, 10 mM DTT；20410-B1：500 mM Tris-HCl, pH 8.0, 2% Igepal (NP-40), 10 mM DTT. |
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Storage
This product should be stored at -85~-65℃ for 1 year.
Instructions
- The following reaction system is formulated in the EP tube:
- Incubate at 30℃, suck out 20 μL of the above reaction solution at 1, 2, 4 and 6 hours in a separate EP tube respectively.
- Add 20 μL of 2×SDS loading Buffer into the EP tube and place it at -20℃.
- After the reaction of all samples was completed, the samples were boiled for 5 min and 30 μL was taken for SDS-PAGE analysis. Determine the optimal response time.
|
Components |
Volume (μL) (The reaction system can be scaled down equally) |
|
Fusion protein |
50 μg |
|
10×Protease Buffer |
20 μL |
|
SUMO Protease (1U/μL) |
10 μL |
|
ddH2O |
to 200 μL |
Note: If the fusion protein requires low temperature treatment, the reaction solution can be placed at 4℃ to prolong the reaction time and increase the amount of SUMO enzyme to ensure the effect of enzyme digestion.
Table 1. SUMO Protease digestion activity at different temperatures
|
Incubation Time（h） |
Digestion activity at different temperatures (%) |
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|
4℃ |
16℃ |
25℃ |
30℃ |
|
|
0.5 |
48 |
73 |
83 |
88 |
|
1 |
60 |
87 |
90 |
93 |
|
2 |
71 |
94 |
94 |
95 |
|
3 |
74 |
95 |
95 |
95 |
Notes
1. Please wear the necessary PPE, such lab coat and gloves, to ensure your health and safety.
2. This product is for research use only.