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SUMO Protease
| No | Product Name | Cat.# | Conc | Size | Price(￦) | Add |
|---|---|---|---|---|---|---|
| 1 | SUMO Protease | PR001S | 1 unit/μl | 250 units | ￦ 300,000 |
|
| 2 | SUMO Protease | PR001M | 1 unit/μl | 500 units | ￦ 540,000 |
|
| 3 | SUMO Protease | PR001L | 1 unit/μl | 1,250 units | ￦ 1,200,000 |
|
SUMO Protease (Ulp1, Ubl-specific protease 1) from yeast is expressed and purified from E. coli to near homogeneity, 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, it cleaves a peptide bond located after the C-terminal diglycine residues of SUMO. The optimal temperature for cleavage is 30oC; however, the enzyme is active over wide ranges of temperature and pH (pH 7.0-9.0).
Characteristics
- Molecular weight : 25 kDa
- Reaction temperature : 2-37℃ (30℃ is optimal)
- Optimal pH range : pH 5.5-9.5
Applications
- Removal of fusion tag from recombinant proteins
Quality control
- Purity : >99% on SDS-PAGE
- Endonuclease-free
- Exonuclease-free
- RNase-free
Unit definition
One unit of SUMO Protease is the amount of enzyme required to cleaves 100 μg of SUMO-MetGFP substrate in 1X SUMO Protease buffer at 30°C for 1 hr.
Storage buffer
50 mM Tris-HCl (pH 8.0), 150 mM NaCl, 5 mM DTT, 4% trehalose, 50% glycerol
10X SUMO Protease buffer
- Salt plus buffer : 500 mM Tris-HCl, pH 8.0, 2% Igepal (NP-40), 1.5 M NaCl, 10 mM DTT
- Salt free buffer : 500 mM Tris-HCl, pH 8.0, 2% Igepal (NP-40), 10 mM DTT
Standard reaction conditions
|
10X SUMO Protease buffer +/-salt |
20 μl |
|
SUMO protease (1 units/μl) |
10 μl |
|
Fusion protein (substrate, 1 μg/μl) |
20 μl |
|
Distilled water |
up to 200 μl |
→ Incubate the reaction mixture at 30℃.
→ Withdraw 20 μl aliquots at 1, 2,
4, and 6 hr after incubation.
→ Add 20 μl of 2X SDS sample buffer
to the aliquots, boil the mixture, and analyze by SDS-PAGE
to check.
→ Keep the rest of the reaction mixture
at -20℃ until analysis is completed
Cautions
- Avoid
repeated cycles of freezing-thawing.
- This enzyme is
active in NaCl from 100 mM to 300 mM (150 mM is optimum).
- Keep the
imidazole concentration lower than 150 mM. Concentrations higher than 150 mM
can inhibit the
activity of this protease.
A SUMO-fusion protein (20 μg), in which SUMO is fused to its Nterminus, was incubated at 4, 16, 21, or 30℃ for 1 hr with 10 unit of SUMO proteases from Enzynomics (Cat.# PR001) or other company (Supplier A). At all temperatures tested, SUMO was released from the substrates by SUMO proteases of both companies. The optimal temperature for SUMO proteases is 30℃ In excess of enzymes, however, all substrates can be cleaved over a wide range of temperatures. Mw, molecular weight marker. Lane 9, substrate only control. The protein substrate used, cleavage products, and SUMO released by digestion are indicated by arrows.
● Material Safety Data Sheet
