Additional Information
Product Name: | Unconjugated (Free) Polyubiquitin Chain Capture Kit |
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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.