Additional Information
| Product Name: | Nedd8 Conjugation Kit |
|---|---|
| Also Known As: | N/A |
| Catalog No.: | J3150 |
| Size: | 1 Kit |
| Molecular Weight: | N/A |
| Species: | Human |
| Source: | Bacterial recombinant |
| Stock: | 20 mM Tris, 150 mM NaCl, 2 mM βME, 10% Glycerol |
| Concentration: | See tube label |
| Quality Assurance: | 95% based on SDS-PAGE |
| Storage: | Store at -80°C; avoid multiple freeze-thaw cycles |
| PDF Data Sheet: | PDF datasheet, MSDS |
| NCBI RefSeq: | N/A |
| Image(s): | No |
| Shipping Method: | Dry ice shipping |
| References: | N/A |
Details
* This kit is sufficient for setting up 25 x 20 µl reactions.
Description
Nedd8 Conjugation Kit is designed for assaying the conjugation of Nedd8 to protein substrates in vitro. In a protein neddylation reaction, Nedd8 is first activated by the Nedd8 activating enzyme APPBP1/UBA3 (E1) in an ATP hydrolysis-dependent manner; activated Nedd8 is then transferred to the Nedd8 conjugating enzyme Ube2M (UbcH12) (E2) by formation of a thiolester bond with the catalytic cysteine residue of UbcH12I; Nedd8-charged Ube2M (UbcH12) can mediate protein neddylation in the presence of an appropriate Nedd8 ligase (E3).
A typical 20 µl reaction contains 100 nM APPBP1/6xHis-UBAs, 2 µM 6xHis-Ube2M (UbcH12), 15 µM 6xHis-Nedd8, 4 mM ATP, 2 µM E3 (not supplied) and 2 µM protein substrate (no supplied) in the Neddylation Buffer.
Components
• 20X APPBP1/6xHis-UBA3 (2 µM)................25 µl
• 20X 6xHis-Ube2M (UbcH12) (40 µM))..........25 µl
• 10X 6xHis-Nedd8 (150 µM)..........................50 µl
• 10X ATP (40 mM)........................................60 µl
• 10X Neddylation Buffer...............................100 µl
