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.