Linear polyUb Chain-binding Protein Identification / Validation Kit Reference: RP10211K This kit is designed for two purposes: 1) identification of proteins that specially bind linear polyUb chains in whole cell or tissue lysates; 2) validation of direct interactions between linear polyUb chains and their binding proteins. The non-cleavable 6xHis-tagged linear Ub6 is resistant of deubiquitination when incubating with whole cell or tissue lysates, thus allowing maximally capture proteins that specifically bind linear polyUb chains. After binding, 6XHis-tagged non-cleavable linear Ub6 and the binding proteins can be enriched with Nickel XPure Agarose Resin (included). The bound proteins and the non-cleavable polyUb chains can be eluted from Ni resin using a buffer containing 250 mM imidazole (included) or by incubating with TEV protease (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.
Immunoproteasome Activity Fluorometirc Assay Kit II Reference: RP10223K Immunoproteasome Activity Fluorometric Assay Kit II was designed for assaying the immunoproteasomes' chymotrypsin-like, trypsin-like and caspase-like activities in vitro using purified proteasomes, cell lysates or tissue extracts. The proteasome cleaves these fluorogenic substrates, and the released AMC fluorescence can be monitored using a plate reader or fluorometer at the excitation/emission wavelength of 360nm/460nm, respectively.?
K29/K33 polyubiquitin Chain Capture Kit Reference: RP10233K This kit is designed for enrichment of cellular proteins conjugated with lysine 29/33-linked polyubiquitin chains using GST-TRABID (ZNF1). Polyubiquitinated proteins bound on GST-TRABID (ZNF1) can be precipi-tated using glutathione resin and eluted by a buffer containing 10 mM glutathione. Enriched polyubiqui-tinated proteins can be assayed by immunoblotting or mass spectrometry.