NEDD8 Conjugation Initiation Kit
- 貨號 Cat. No.
- K-800
- 規格 Size
- 1 kit
- 品牌 Brand
- Boston Biochem
Description
NEDD8 Conjugation Initiation Kit
human recombinant
Product Information
1 kit
Use & Storage
Store at -80°C. Avoid multiple freeze/thaw cycles.
Catalog #: K-800
NEDD8 Conjugation Initiation Kit
Product Source:
human recombinant
This kit is designed for the conjugation of the ubiquitin-like modifier NEDD8 to proteinsubstrates in vitro, which requires the activities of the heterodimeric human E1 activating enzyme(APPBP1/Uba3) and the UbcH12 E2 enzyme. The E1enzyme charges the NEDD8 by forming an ATP-dependent high energy thiolester bond with the active site cysteine of Uba3. The activated NEDD8 is subsequently transferred to UbcH12and this E2-S-Ub thiolester complex can be used for the conjugation of NEDD8 to protein substrates in the presence of the appropriate E3 enzymes (not supplied).
NOTE: Kit contains reagents sufficient for 10 x 20 μl reactions.
Product Information
Quantity:
1 kit
Supplied:
| Protein | Concentration | Quantity |
| 10X NEDD8 E1 Enzyme | X mg/ml (X μM) | 20 μL |
| 10X NEDD8 | X mg/ml (X μM) | 20 μL |
| 10X UbcH12 | X mg/ml (X μM) | 20 μL |
| 10X Mg-ATP Solution | X mM | 20 μL |
| 10X Reaction Buffer | X mM Hepes pH 8 | 20 μL |
| X mM NaCl | ||
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.
Use & Storage
Storage:
Store at -80°C. Avoid multiple freeze/thaw cycles.