SK-20

Material Data Sheet

NEDD8 Conjugation Rxn Buffer Kit

Price: $59.00
Catalog #: SK-20

NEDD8 Conjugation Rxn Buffer Kit

Price: 
$59
Quantity: 
1 kit

NEDD8 reaction buffer kit contains the optimal buffer formulations for use in assays for the conjugation of the ubiquitin-like modifier NEDD8 (UL-812) to protein substrates in vitro. This requires the activities of the heterodimeric human E1 activating enzyme (APPBP1/Uba3) (E-313) and the UbcH12 E2 enzyme (E2-656). This buffer can be used to supplement NEDD8 conjugation kit (K-800).

Product Information

Stock: 
1. 1ml 10X NEDD8 Reaction Buffer2. 100μl 10X Mg-ATP3. 1ml 100X E1 Stop Buffer4. 1ml 5X Non-reducing gel loading buffer
Background: 
The ubiquitin-like NEDD8 is conjugated to a variety of proteins in the presence of UbcH12 and an E1 activating enzyme. The NEDD8 E1 enzyme uses ATP to adenylate the C-terminal glycine residue of NEDD8, forming a high-energy thiolester bond. The second step is the trans-esterification reaction whereby the activated NEDD8 is transferred to the active site cysteine of UbcH12. UbcH12 is a<br>member of the E2 family and is homologous to ubiquitin-conjugating enzymes, but is specific for the conjugation of NEDD8 to a variety of target proteins. Nedd8 plays a critical regulatory role in cell proliferation and development, and modifies nearly all members of the Cullin family.
Storage: 

Store at -20°C. Avoid multiple freeze/thaw cycles.

Literature

References: 

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Morimoto M., et al. (2003) Biophys. Biochem. Res. Comm. 301:392-398
Wada H., et al. (1999) Biophys. Biochem. Res. Comm. 275:100-105
Whitby F.G.,et al. (1998) J. Biol. Chem. 273: 34983-34991