A useful reagent for studying SUMOylation in vitro. Represents a readily detectable alternative to radio-iodinated SUMO-3. Biotinylated SUMO-3 is activated by SUMO E1, a competent substrate for the SUMO E2 conjugating enzyme Ubc9. Can be conjugated to target proteins, in either a SUMO E3 ligase dependent or independent manner, as appropriate. The biotinyl moiety also provides a potential route to affinity purification of conjugated substrates (or thiol ester-linked enzymes) on avidin/streptavidin matrices.
Product Specification
| Alternative Name: | Sentrin-3, Small ubiquitin-related modifier 3, SUMO-2 |
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| MW: | ~13kDa (based on Bt3-SUMO-2 as predominant form present). |
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| Source: | Produced in E. coli. |
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| UniProt ID: | P55854 |
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| Formulation: | Lyophilized solid (from 1:1 acetonitrile:water). |
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| Application Notes: | Suggested uses:- Ready detection of SUMO-3 modification of specific proteins in vitro.
- Demonstration that novel proteins are potential targets for SUMOylation under in vitro conditions.
- Purification of SUMO-3 conjugates via streptavidin affinity chromatography.
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| Reconstitution: | In order to ensure full solubility it is recommended that the material be dissolved in 5µL dimethyl sulphoxide (DMSO) prior to dilution with buffer of choice. |
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| Shipping: | Shipped on Blue Ice |
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| Short Term Storage: | -20°C |
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| Long Term Storage: | -20°C |
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| Use/Stability: | Stable for at least 12 months after receipt when stored at -20°C. |
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| Handling: | Avoid freeze/thaw cycles. |
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| Scientific Background: | SUMO (small ubiquitin-related modifier) is a member of a family of ubiquitin-like proteins involved in the regulation of the cellular functions of a wide variety of proteins. Four members of the SUMO family have been described in vertebrates, SUMO-1 and the close homologues SUMO-2 and SUMO-3, with some 50% homology between SUMO-1 and SUMO-2/3. |
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| Technical Info/Product Notes: | Note: SUMO-3 contains 8 lysine residues, any/many of which are likely to be biotinylated in this product. These modification sites have not been mapped. |
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| Regulatory Status: | RUO - Research Use Only |
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Western blot of biotinylated-SUMO-3 (
BML-UW0555) thioester assay. Bt-SUMO3 conjugated to Ubc9 (
BML-UW9320) in the presence of SUMO E1 (
BML-UW9330). Bt-SUMO-3 conjugated species detected with streptavidin-HRP."
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General Literature References
Sumoylation of heterogeneous nuclear ribonucleoproteins, zinc finger proteins, and nuclear pore complex proteins: a proteomic analysis: T. Li, et al.; PNAS
101, 8551 (2004),
Abstract;
In vitro SUMO-1 modification requires two enzymatic steps, E1 and E2: T. Okuma, et al.; BBRC
254, 693 (1999),
Abstract;
