Additional Information
| Product Name: | Sumo3 Conjugation Kit |
|---|---|
| Also Known As: | N/A |
| Catalog No.: | J3130 |
| 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
Sumo3 Conjugation Kit is designed for assaying the conjugation of Sumo3 to protein substrates in vitro. In a protein sumoylation reaction, a Sumo protein is first activated by the Sumo activating enzyme SAE1/SAE2 (E1) in an ATP hydrolysis-dependent manner; activated Sumo is then transferred to the Sumo conjugating enzyme Ube2I (E2) by formation of a thiolester bond with the catalytic cysteine residue of Ube2I; Sumo-charged Ube2I can mediate protein sumoylation in the presence of an appropriate Sumo ligase (E3).
A typical 20 µl reaction contains 100 nM SAE1/SAE2, 2 µM 6xHis-Ube2I, 50 µM Sumo3, 4 mM ATP, 2µM E3 (not supplied) and 2 µM protein substrate (no supplied) in the Sumoylation Buffer.
Components
• 20X SAE1/SAE2 (2 µM)..........................25 µl
• 20X 6xHis-Ube2I (40 µM)........................25 µl
• 10X Sumo3 (500 µM)..............................50 µl
• 10X ATP (40 mM)....................................60 µl
• 10X Sumoylation Buffer.........................100 µl
