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Recombinant Human NEDD8 AMC is a fluorogenic substrate for some NEDD8-specific isopeptidases. Release of AMC fluorescence can be monitored with an excitation wavelength of345 nm and an emission wavelength of 445 nm. Reaction conditions will need to be optimized for each specific application. We recommend an initial Recombinant Human NEDD8 AMC concentration of 0.1-1 μM.

Source

E. coli-derived human NEDD8 protein

Accession #

Q15843

Predicted Molecular Mass

8.7 kDa

Product Datasheets

Product Datasheet

COA

SDS

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins.Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration.The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard.In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

UL-552

Formulation	X mg/ml (X μM) in 50 mM HEPES pH 7.5,100 mM NaCl
Shipping	The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage:	Protect from light. Use a manual defrost freezer and avoid repeated freeze-thaw cycles. 12 months from date of receipt, -70 °C as supplied. 3 months, -70 °C under sterile conditions after opening.

Reconstitution Calculator

Background: NEDD8

Neural Precursor Cell Expressed Developmentally Downregulated Gene 8 (NEDD8), also known as Related to Ubiquitin 1 (Rub1), is a 6-8 kDa member of the Ubiquitin family of proteins. Human pro-NEDD8 is 81 amino acids (aa) in length. Pro-NEDD8 contains one Ubiquitin-like domain (aa 1-65) and a 4-5 aa C-terminal propeptide (1,2). Cleavage of the propeptide exposes a C-terminal glycine residue that is used to generate a glycine-lysine intermolecular bond. Mature human NEDD8 shows 100% aa identity to NEDD8 from mouse, rat, and canine. Human NEDD8 is activated by a distinct NEDD8-activating (E1) enzyme, a heterodimeric complex composed of APPBP1 and UBA3 subunits (3). Activated NEDD8 is subsequently transferred to the UBE2M/Ubc12 or UBE2F NEDD8-conjugating (E2) enzymes (4). Through a process termed neddylation, the ROC1/Rbx1 RING Finger E3 ligase transfers NEDD8 to specific substrates (5). For example, transfer of NEDD8 to the Cullin scaffold activates members of the Cullin-RING Ligase (CRL) family of Ubiquitin ligases (E3) (6-8). CRL-induced ubiquitination is known to regulate proteins that are important for cell cycle progression and cell survival (9,10). Physiologically, NEDD8 plays a critical regulatory role in cell proliferation and dysregulation of the NEDD8 pathway has been associated with several cancer pathologies (11,12).

References

Kumar, S. et al. (1993) Biochem. Biophys. Res. Commun. 195:393.
Kamitani, T. et al. (1997) J. Biol. Chem. 272:28557.
Huang, D.T. & B.A. Schulman (2005) Methods Enzymol. 398:9.
Gong, L. & E.T. Yeh (1999) J. Biol. Chem. 274:12036.
Ohki, Y. et al. (2009) Biochem. Biophys. Res. Commun. 381:443.
Petroski, M.D. & R.T. Deshares (2005) Nat. Rev. Mol. Cell Biol. 6:9.
Hjerpe, R. et al. (2012) Biochem. J. 441:927.
Calabrese, M.F. et al. (2011) Nat. Struct. Mol. Biol. 18:947.
Tateishi, K. et al. (2001) J. Cell. Biol. 155:571.
Ohh, M. et al. (2002) EMBO Rep. 3:177.
Luo, Z. et al. (2012) Cancer Res. 72:3360.
Soucy, T.A. et al. (2010) Genes Cancer 1:708.

Long Name

Neural Precursor Cell Expressed Developmentally Down-Regulated Protein 8

Entrez Gene IDs

4738 (Human); 18002 (Mouse); 25490 (Rat)

Alternate Names

FLJ43224; MGC104393; MGC125896; MGC125897; NEDD8; NEDD-8; Neddylin; Neural precursor cell expressed developmentally down-regulated protein 8; neural precursor cell expressed, developmentally down-regulated 8; Rub1; Ubiquitin-like protein Nedd8

Related Research Areas

Developmental Factors Implicated in Neurodegeneration
Ubiquitin-like Modifiers (UBLs) and Regulators

Citations for Recombinant Human NEDD8 AMC Protein, CF

R&D Systems personnel manually curate a database that contains references using R&D Systems products.The data collected includes not only links to publications in PubMed,but also provides information about sample types, species, and experimental conditions.

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Crystal structure and activity-based labeling reveal the mechanisms for linkage-specific substrate recognition by deubiquitinase USP9XAuthors: P Paudel, Q Zhang, C Leung, HC Greenberg, Y Guo, YH Chern, A Dong, Y Li, M Vedadi, Z Zhuang, Y TongProc. Natl. Acad. Sci. U.S.A., 2019;0(0):.Applications: Bioassay
Nedd8 hydrolysis by UCH proteases in Plasmodium parasitesAuthors: M Karpiyevic, S Adjalley, M Mol, DB Ascher, B Mason, GJ van der He, H Laman, H Ovaa, MCS Lee, K Artavanis-PLoS Pathog., 2019;15(10):e1008086.Species: HumanSample Types: Whole CellsApplications: Bioassay
A Wolbachia deubiquitylating enzyme induces cytoplasmic incompatibilityAuthors: JF Beckmann, JA Ronau, M HochstrassNat Microbiol, 2017;2(0):17007.Species: N/ASample Types: ProteinApplications: Bioassay
Diverse mechanisms of metaeffector activity in an intracellular bacterial pathogen, Legionella pneumophilaAuthors: Malene L UrbanusMol. Syst. Biol, 2016;12(12):893.Applications: Bioassay
Structural and biochemical characterization of the Cop9 signalosome CSN5/CSN6heterodimer.Authors: Birol M, Enchev R, Padilla A, Stengel F, Aebersold R, Betzi S, Yang Y, Hoh F, Peter M, Dumas C, Echalier APLoS ONE, 2014;9(8):e105688.Species: N/ASample Types: ProteinApplications: Bioassay