- Regulatory Status
- RUO
- Other Names
- Small Ubiquitin Like Modifier 3, Small Ubiquitin-Related Modifier 3, SMT3H1, SUMO-3, SMT3A
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The ability of recombinant human SUMO3-GFP protease substrate to be cleaved by different proteases (SENP2, thrombin, ENTK, 3C, factor Xa, and TEV) is visible by an SDS-PAGE gel. -
Stability Testing for Recombinant Human SUMO3-GFP Protease Substrate. Human SUMO3-GFP was aliquoted in 20 mM Tris pH 8.0, 100 mM NaCl. One aliquot was frozen and thawed four times (4x Freeze/Thaw) and compared to the control that was kept at 4°C (Control). The samples were tested for their ability to be cleaved by different proteases (3C and thrombin). Cleavage of human SUMO3-GFP is visible by an SDS-PAGE gel.
| Cat # | Size | Price | Quantity Check Availability | ||
|---|---|---|---|---|---|
| 793304 | 25 µg | $118.00 | |||
| 793306 | 100 µg | $211.00 | |||
Select size of product is eligible for a 40% discount! Promotion valid until December 31, 2024. Exclusions apply. To view full promotion terms and conditions or to contact your local BioLegend representative to receive a quote, visit our webpage.
The protein, SUMO3, is a member of the small ubiquitin related modifier (SUMO) family. At least three SUMO family members, SUMO1, SUMO2, and SUMO3, are found in mammals. SUMO3 shares a 42% sequence identity with SUMO1 and a 96% identity with SUMO2. After translation, the protein is modified and covalently conjugates to other proteins. This post-translation modification is referred to as sumolyation. SUMO proteins are linked to other proteins via an isopeptide bond formed between their carboxyl-terminal glycine residues and the lysine(s) of the substrate. Most of the substrate proteins that SUMO modifies are located in the nucleus. The mechanism of conjugation of SUMO with substrates parallels to the mechanism of ubiquitination. Sumolyation is integrated in many cellular processes such as DNA replication and repair, nuclear transport, signal transduction, and regulation of transcription. SUMO3 is successfully cleaved by different proteases. A family of sentrin/SUMO-specific proteases (SENPs) regulates the reversible process of sumolyation. SENP2 is a protease that de-modifies SUMO3 and other SUMO family protein conjugates. SUMO3-GFP is successfully cleaved by the following enzymes: SENP2, thrombin, enterokinase cleavage enzyme (EKT), active HRV14 3C, TEV, and factor Xa. SUMO fusion technology has been use to enhance protein expression, solubility, and purification in prokaryotes. E.coli lacks the SUMO protease, therefore the purification of SUMO-tag proteins is achievable. A different fusion system called split SUMO has been developed for the expression of SUMO-tagged proteins in eukaryotic systems.
Product Details
- Source
- Human SUMO3, amino acid (Asn14-Gly92) (Accession # P55854.2) was expressed in pBL-2 cells. The N terminal has a Met and 6x His tag, followed by different protease cleavage [SENP2, thrombin, enterokinase, 3C protease, factor Xa, and tobacco Etch virus (TEV)] and C-terminal GFP (AAB02574.1).
- Molecular Mass
- The 356 amino acid recombinant protein has a predicted molecular mass of approximately 40.4 kD. The DTT-reduced and non-reduced protein migrates at approximately 50 kD by SDS-PAGE. The predicted N-terminal amino acid is Met.
- Purity
- > 95%, as determined by Coomassie stained SDS-PAGE
- Formulation
- 0.2 µm filtered protein solution is in 20 mM Tris pH 8.0, 100 mM NaCl.
- Endotoxin Level
- Less than 0.1 EU per µg cytokine as determined by the LAL method
- Concentration
- 25 µg size is bottled at 200 µg/mL. 100 µg size and larger sizes are lot-specific and bottled at the concentration indicated on the vial. To obtain lot-specific concentration and expiration, please enter the lot number in our Certificate of Analysis online tool.
- Storage & Handling
- Unopened vial can be stored between 2°C and 8°C for up to 2 weeks at -20°C for up to six months, or at -70°C or colder until the expiration date. For maximum results, quick spin vial prior to opening. The protein can be aliquoted and stored at -20°C or colder. Stock solutions can also be prepared at 50 - 100 µg/mL in appropriate sterile buffer, carrier protein such as 0.2 - 1% BSA or HSA can be added when preparing the stock solution. Aliquots can be stored between 2°C and 8°C for up to one week and stored at -20°C or colder for up to 3 months. Avoid repeated freeze/thaw cycles.
- Activity
- Human SUMO3-GFP protease substrate is cleaved successfully by SENP2, thrombin, enterokinase, 3C protease, factor Xa, and tobacco Etch virus (TEV).
- Application
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Bioassay
- Application Notes
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Human SUMO3-GFP Protocol:
Human SUMO3-GFP is digested by different proteases.
- Incubate mixtures of 5 µg of SUMO3-GFP with protease (eigther SENP2, thrombin, enterokinase (ENTK), 3C protease, factor Xa, TEV) for 16 hours at 25°C.
- Run SDS-PAGE gel with mixtures.
Assay Buffer for Each Protease Mixture:
a) Human SENP2 and 3C protease buffer: 20 mM Tris, pH8, 100 mM NaCl
b) Thrombin and factor Xa buffer: 20 mM Tris, pH 8, 100 mM NaCl, 2 mM CaCl2
c) Enterokinese buffer: 20 mM Tris, pH 8, 100 mM NaCl
d) TEV buffer: 20 mM Tris, pH 8, 100 mM NaCl, 1 mM DTT, 0.5 mM EDTA
BioLegend carrier-free recombinant proteins provided in liquid format are shipped on blue-ice. Our comparison testing data indicates that when handled and stored as recommended, the liquid format has equal or better stability and shelf-life compared to commercially available lyophilized proteins after reconstitution. Our liquid proteins are validated in-house to maintain activity after shipping on blue ice and are backed by our 100% satisfaction guarantee. If you have any concerns, contact us at tech@biolegend.com. - Additional Product Notes
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Antigen Details
- Structure
- Monomer
- Distribution
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SUMO is ubiquitously expressed.
- Function
- Functions in post-translational modification system
- Ligand/Receptor
- Substrate for: SEMP2, thrombin, enterokinase, 3C protease, factor Xa and TEV
- Bioactivity
- SUMO3-GFP protease substrate is cleaved successfully by different proteases including: SENP2, thrombin, enterokinase, prescission 3C, factor Xa and TEV.
- Cell Type
- B cells, Epithelial cells, Neurons, T cells
- Biology Area
- Cardiovascular Biology, Cell Biology, Cell Cycle/DNA Replication, DNA Repair/Replication, Neuroscience
- Molecular Family
- Proteases
- Antigen References
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- Saitoh H, et al. 1997. Trends Biochem Sci. 22:374.
- Yeh ETH, et al. 2000. Gene. 248:1.
- Zhang H, et al. 2002. Mol Cell Biol. 22:6498.
- Reverter D, et al. 2004. Structure. 12:1519.
- Butt TR, et al. 2005. Prot Expr Purif. 43:1.
- Yeh ETH, et al. 2006. J Biol Chem. 281:15869.
- Gene ID
- 6612 View all products for this Gene ID
- UniProt
- View information about SUMO3 on UniProt.org