SK-20

Material Data Sheet

NEDD8 Conjugation Reaction Buffer Kit

Price: $50.00
Catalog #: SK-20

NEDD8 Conjugation Reaction Buffer Kit

Price: 
$50
Quantity: 
1 kit
Data Sheet: 
application/pdf iconSK-20_NEDD8 Conjugation Reaction Buffer Kit_.pdf
Stock: 

1. 1ml 10X NEDD8 Reaction Buffer
2. 100μl 10X Mg-ATP
3. 1ml 50X E1 Stop Buffer
4. 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 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.

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).

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

Literature

References: 

Gong L., et al. (1999) J. Biol. Chem. 274: 12036-12042
Hori T., et al. (1999) Oncogene. 18:6829-6834
Kamura T., et al. (1999) Genes. Dev. 13:2928-2933
Kumar S., et al. (1993) Biophys. Biochem. Res. Comm. 195:393-399
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