Recombinant SENP2 (His-tag)

Item Number

R752159

• Product Name: Recombinant SENP2 (His-tag)
• Grade: EnzymoPure™
• Specifications & Purity: ≥95%(SDS-PAGE), other protease activity not detected.

Product Description

Aladdin's Recombinant SENP2 (His-tag), also named as rSENP2 (His-tag), is a protease containing the activity domain (Asp364-Leu589) of human SENP2. It is recombinantly expressed in E. coli, with a His tag and a theoretical molecular weight of 26.8 kDa. It is commonly used for in vitro DeSUMOylation or excision of SUMO1, SUMO2 or SUMO3 tags from recombinant fusion proteins.Instead of recognizing a specific amino acid sequence, rSENP2 specifically recognizes the three-dimensional structures of human SUMO1/2/3 and efficiently cleaves the QTGG/X amino acid sequence at their termini to release the mature form of SUMO1/2/3.This product is fused with a N-terminal His tag. It can be detected with the specific antibody or be removed using a nickel column. Small ubiquitin-like modifier (SUMO) is a protein family ubiquitous in eukaryotes. There are four classes of SUMO in human

Precautions：

This product contains 50% glycerol and will not freeze at -20°C. Storage at -80°C should be avoided as freeze-thaws may reduce the enzymatic activity of this product.This product is viscous. Make sure the exact amount is taken. Mix after adding it to solutions by gentle pipetting and avoid the formation of air bubbles.The enzymatic activity of this product is largely impacted by the spatial structure of proteins fused with SUMO tag. For cases where the enzymatic activity is relatively low, the amount of enzyme should be increased significantly. To obtain higher enzymatic activity, manipulating the amino acid sequence between SUMO tag and target protein can be attempted.This product is for R&D only. Not for drug, household, or other uses.For your safety and health, please wear a lab coat and disposable gloves during the operation.

Instructions for Use：

1. Optimization of digestion conditions. The ratio of enzyme to tagged protein should be optimized for different SUMO3-containing proteins as follows:a. Add 1μl of rSENP2 (100U/μl) to 99μl of Reaction Buffer and mix well. Reaction Buffer is not supplied in this product.Note: As rSENP2 is highly active over a wide pH (6.0-10.0), temperature (2-37ºC), and ionic strength (0-1M NaCl, 0-500mM Imidazole), there is no specific restrictions on the components of Reaction Buffer. The enzymatic activity of rSENP2 is higher in the presence of 0.5-2mM DTT, so the addition of appropriate amount of DTT to the reaction can be considered for experimental needs. b. Set up the reaction in a 1.5ml centrifuge tube as follows:ComponentVolume SUMO3-tag Protein (10μg) xμl rSENP2 (1U/μl) 0, 1, 5 or 10μl Reaction Buffer To 20μl c. Incubate the reaction. Please refer to the following table for incubation temperature and time.Temperature Incubation Time 4℃ overnight 16℃ 4h 25℃ 2h 37℃ 1h d. Add 4μl of SDS-PAGE Sample Loading Buffer (6X) ( Coomassie Blue Super Fast Staining Solution (, P0017F).e. Observe proteins in the SDS-PAGE gel. The ideal amount of rSENP2 is that can just completely excise the SUMO3 tag. The reaction system can be scaled up or down proportionally.f. If the SUMO3 tag is not completely excised, increasing the amount of rSENP2, the digestion time or the reaction temperature can be attempted.2. Enzymatic digestion and purification. With the optimal digestion conditions, the reaction system can be scaled up to remove the SUMO3 tag from the target protein. After desumoylation, the excised SUMO3 tag with His tag and the rSENP2 with His tag can be removed by the nickel column, and the high-purity target protein without SUMO3 tag can be obtained. It is also possible to perform on-column digestion of fusion proteins with both His-tag and SUMO3 tag. If the capacity of the nickel column is sufficient, both rSENP2 (His-tag) and the cleaved SUMO3 tag with His-tag will be bound to the nickel column while only the target protein will be eluted.