The non-cleavable 6xHis-tagged K63 polyUb chains (2-4) are resistant of deubiquitination when incubating with whole cell or tissue lysates, thus allowing maximally capture proteins that specifically bind K63 polyUb chains. After binding, the non-cleavable K63 polyUb chains and the binding proteins can be enriched by Nickel XPure Agarose Resin (included). The bound proteins and the non-cleavable K63 polyUb chains can be eluted from Ni resin using a buffer containing 250 mM imidazole (included) or by incubating with thrombin (not included). Either of the elution method can significantly reduce the amounts of non-specific binding proteins when compared with elution using SDS sample buffer.
Additional Information
| Product Name: | K63 polyUb Chain-binding Protein Identification / Validation Kit |
|---|---|
| Also Known As: | N/A |
| Catalog No.: | J2510 |
| 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: | See the storage condition on the label of each component; avoid multiple freeze-thaw cycles |
| PDF Data Sheet: | PDF datasheet |
| NCBI RefSeq: | N/A |
| Image(s): | No |
| Shipping Method: | Dry ice shipping |
| References: | N/A |
Details
* This kit is designed for two purposes: 1) identification of proteins that specially bind K63 polyUb chains in whole cell or tissue lysates; 2) validation of direct interactions between K63 polyUb chains and their binding proteins.
Description
The non-cleavable 6xHis-tagged K63 polyUb chains (2-4) are resistant of deubiquitination when incubating with whole cell or tissue lysates, thus allowing maximally capture proteins that specifically bind K63 polyUb chains. After binding, the non-cleavable K63 polyUb chains and the binding proteins can be enriched by Nickel XPure Agarose Resin (included). The bound proteins and the non-cleavable K63 polyUb chains can be eluted from Ni resin using a buffer containing 250 mM imidazole (included) or by incubating with thrombin (not included). Either of the elution method can significantly reduce the amounts of non-specific binding proteins when compared with elution using SDS sample buffer.
Components
• 6xHis-Non-cleavable K63 polyUb chain (2-4) [Store at -80°C]..........100 µg
• 6xHis-Ubiquitin [Store at -80°C].............................................................................100 µg
• Nickel XPure Agarose Resin (suspended in 50% slurry) [Store at 2-8°C]....................500 µl
• 4 M Imidazole [Store at -80°C]......................................................................................500 µl
• Iodoacetamide( IAA ) [Store at -80°C]..........................................................................25 mg
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