Product Details
| MW: | ~11kDa (based on Bt3-NEDD8 as predominant form present). |
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| Source: | Produced in E. coli. |
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| UniProt ID: | Q15843 |
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| Formulation: | Lyophilized. Contains PBS. |
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| Reconstitution: | Reconstitute with sterile distilled water. |
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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: | Biotinylated NEDD8 is a useful reagent for studying NEDDylation in vitro, where it represents a readily detectable alternative to radio-iodinated NEDD8. Biotinylated NEDD8 is activated by NEDD8 E1, is a competent substrate for the NEDD8 E2 conjugating enzyme UbcH12 and can be conjugated to target proteins in the presence of a suitable NEDD8 E3 ligase. The biotinyl moiety also provides a potential route to affinity purification of conjugated substrates (or thiol ester-linked enzymes) on avidin/streptavidin matrices. NEDD8 contains 9 lysine residues, any/many of which are likely to be biotinylated in this product. These modification sites have not been mapped. Biotinylated NEDD8 is prepared from recombinant human His-tagged NEDD8 (Prod. No. BML-UW9225). |
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| Technical Info/Product Notes: | Biotinylated mature protein for NEDD8 conjugation and NEDDylation pathway studies. Suggested uses: - Ready detection of NEDD8 modification of specific proteins in vitro.
- Demonstration that novel proteins are potential targets for NEDDylation under in vitro conditions.
- Purification of NEDD8-conjugates via streptavidin affinity chromatography.
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| Regulatory Status: | RUO - Research Use Only |
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Western blot of biotinylated-NEDD8 (Prod. No. BML-UW0560) thioester assay. Bt-NEDD8 conjugated to UbcH12 (Prod. No.
BML-UW9145) in the presence of NEDD8 E1 (Prod. No.
ENZ-PRT112). Bt-NEDD8 conjugated species detected with streptavidin-HRP."
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General Literature References
In vitro systems for NEDD8 conjugation by Ubc12: T. Chiba; Methods Enzymol.
398, 68 (2005),
Abstract;
Mdm2-mediated NEDD8 conjugation of p53 inhibits its transcriptional activity: D.P. Xirodimas, et al.; Cell
118, 83 (2004),
Abstract;
A novel mammalian Smt3-specific isopeptidase 1 (SMT3IP1) localized in the nucleolus at interphase: T. Nishida, et al.; Eur. J. Biochem.
267, 6423 (2000),
Abstract;
Identification of the activating and conjugating enzymes of the NEDD8 conjugation pathway: L. Gong & E.T. Yeh; J. Biol. Chem.
274, 12036 (1999),
Abstract;
