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Cat. No.: SEUH-500 (for 500U)

Cat. No.: SEUH-2k (for 2KU)

Cat. No.: SEUH-10k (for 10KU)

Cat. No.: SEUH-50k (for 50KU)

Description

Recombinant SENPEUH (His-tag), also known as rSENPEUH (His-tag), is a newly engineered SENP protease that is expressed through recombinant technology in E. coli. It has a theoretical molecular weight of 25.7 kDa and is primarily used for removing the SUMOEU1 tag from recombinant proteins fused in yeast or eukaryotic cell systems.

The SUMOEU1 tag, a mutant form of SUMO, can enhance the solubility and stability of the target protein in yeast and eukaryotic cells. Moreover, the fusion of the SUMOEU1 tag to the target protein does not participate in SUMO-related cellular regulation and is not cleaved by endogenous desumoylating enzymes. Therefore, the SUMOEU1 tag is suitable for the expression and purification of recombinant proteins in yeast and eukaryotic cells. rSENPEUH specifically recognizes and cleaves the SUMOEU1 tag, enabling efficient separation of the SUMOEU1 tag from the target protein. The cleaved target protein does not contain any residual amino acids from the corresponding tag.

This product is equipped with an N-terminal His-tag, which can be removed by using appropriate His antibodies, resin beads, or nickel columns for affinity purification.

Protein Information

• Name: Recombinant SENPEUH Protease (His-tag)
• Synonyms: SENP1EU, SENPEU, rSENPEUH
• Molecular Weight: 25.7kDa
• Physical Appearance: Liquid
• Biological Activity: 10U/μl
• Unit Definition: One unit is defined as the amount of enzyme required to digest 10µg of SUMOEU1-MBP protein in 1 hour at 25℃, to the extent that the accumulation of SUMOEU1 tag and MBP as determined by SDS-PAGE.
• Purity: ≥95% by SDS-PAGE, other protease activity not detected.
• Storage Buffer: 5mM Tris (pH7.4), 250mM NaCl, 5mM DTT, 50% Glycerol
• Applications: Removal of SUMOEU1 tag from recombinant proteins expressed in yeast or mammalian cell systems.

rSENPEUH exhibits optimal enzymatic cleavage activity at 25℃, within a broad pH range (6.0-10.0), wide temperature range (2-37℃), and broad range of ion strength (0-1M NaCl, 0-500mM Imidazole). For practical purposes, it is recommended to perform overnight enzyme cleavage at 4℃ to maintain the activity of the recombinant protein as much as possible. The enzymatic activity of rSENPEUH is higher in the presence of 0.5-2mM DTT, thus it is advisable to add an appropriate concentration of DTT to the cleavage system to enhance cleavage efficiency.

rSENPEUH's enzymatic activity is not inhibited by commonly used serine protease inhibitors such as PMSF, AEBSF, Bestatin, Pepstatin, E-4, TLCK, and EDTA. However, proteinase inhibitors targeting cysteine residues, such as NEM or IAA, can significantly inhibit the enzymatic activity of rSENPEUH.

When using this product for enzymatic cleavage of recombinant proteins with SUMOEU1 tags, it is recommended to use 1μl of rSENPEUH (10U/μl) for every 100μg of SUMOEU1-tagged fusion protein and perform overnight cleavage at 4℃. The 500U, 2KU, 10KU, and 50KU packaging sizes of this product can be used for approximately 5mg, 20mg, 100mg, and 500mg of SUMOEU1-tagged fusion proteins, respectively.

Precautions

• This product contains 50% glycerol and will not freeze at -20℃. Avoid storing at -80℃ as freeze-thaw cycles may reduce the enzymatic activity of this product.
• The product has a viscous consistency, so it is important to accurately measure the amount when pipetting. After adding the product, ensure thorough mixing by pipetting up and down to avoid bubble formation.
• The enzymatic activity of this product is closely related to the spatial structure of the fusion protein formed by the SUMOEU1 tag and the target protein. While the activity of some SUMOEU1-tagged fusion proteins may reach the activity specified in the product manual, there may be significant differences in the enzymatic cleavage activity for certain SUMOEU1-tagged fusion proteins. If the enzymatic activity of the product is relatively low, a substantial increase in the enzyme dosage is required. If a higher cleavage activity is desired, it is recommended to modify or change the amino acid sequence at the junction between the target protein and the SUMOEU1 tag within permissible limits to achieve better cleavage results.
• This product is intended for scientific research purposes by professionals only and should not be used for clinical diagnosis or treatment, food or drug purposes, or stored in regular household settings.
• For your safety and health, please wear appropriate laboratory attire and disposable gloves when handling this product.

Storage

The minimum shelf life is 2 years at -20°C.

Only for research and not intended for treatment of humans or animals

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