K63 polyUb Chain-binding Protein Identification / Validation Kit

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.

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