Additional Information
| Product Name: | Unconjugated (Free) Polyubiquitin Chain Capture Kit |
|---|---|
| Also Known As: | N/A |
| Catalog No.: | J4420 |
| Size: | 1 Kit |
| Molecular Weight: | No |
| Species: | No |
| Source: | No |
| Stock: | Enzyme stocked in 20 mM Tris, 150 mM NaCl, 2 mM βME, 10% Glycerol |
| Concentration: | See tube label |
| Quality Assurance: | Enzyme >95% by 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: | No |
Details
This kit is sufficient for purification of free polyubiquitin chains in 100 mg or 20 X 5 mg cell or tissue extracts at a concentration of 5 mg/ml.
Description
The Unconjugated (Free) Polyubiquitin Chain Capture Kity enrich free polyUb chains in cell or tissue extracts within 6 hours. This approach uses the N-terminal ZnF-UBP domain of the deubiquitinating enzyme Usp5 to specifically capture free polyUb chains. The ZnF-UBP domain of Usp5 binds the C-terminal diglycine motif in free Ub chains, it does not bind polyUb chains conjugated on substrate proteins.
Components
• GST-ZnF-UBP (4 mg/ml)..............................................0.5 ml
• GST (4 mg/ml).............................................................0.5 ml
• Iodoacetamine (IAA)....................................................25 mg
• Glutathione (reduced).................................................100 mg
• Glutathione Agarose Resin (50% slurry).....................1.5 ml
Images
(Click image to enlarge)
10 µl GST-bound or GST-ZnF-UBP-bound proteins were separated on SDS-PAGEs and immunoblotted with antibodies again Ub or K48-linked polyUb chains.
