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NEDD8 (human), (recombinant) (biotin conjugate)

 
BML-UW0560-0100 100 µg 209.00 USD
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Product Specification

MW:~11kDa (based on Bt3-NEDD8 as predominant form present).
 
Source:Produced in E. coli.
 
UniProt ID:Q15843
 
Formulation:Lyophilized. Contains PBS.
 
Reconstitution:Reconstitute with sterile distilled water.
 
Shipping:Shipped on Blue Ice
 
Short Term Storage:-20°C
 
Long Term Storage:-20°C
 
Use/Stability:Stable for at least 12 months after receipt when stored at -20°C.
 
Handling:Avoid freeze/thaw cycles.
 
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).
 
Technical Info/Product Notes:Biotinylated mature protein for NEDD8 conjugation and NEDDylation pathway studies. Suggested uses:
  1. Ready detection of NEDD8 modification of specific proteins in vitro.
  2. Demonstration that novel proteins are potential targets for NEDDylation under in vitro conditions.
  3. Purification of NEDD8-conjugates via streptavidin affinity chromatography.
 
bml-uw0560 wb
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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bml-uw0560 wb

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;

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