K48 polyUb Chain-binding Protein Identification / Validation Kit Reference: RP10209K This kit is designed for two purposes: 1) identification of proteins that specially bind K48 polyUb chains in whole cell or tissue lysates; 2) validation of direct interactions between K48 polyUb chains and their binding proteins. The Non-cleavable K48 polyUb chain (2-4) are resistant of deubiquitination when incubating with whole cell or tissue lysates, thus allowing maximally capture proteins that specifically bind K48 polyUb chains. After binding, 6xHis-Non-cleavable K48 polyUb chains and the binding proteins can be enriched by Nickel XPure Agarose Resin (included). The bound proteins and the non-cleavable K48 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.
K63 polyUb Chain-binding Protein Identification / Validation Kit Reference: RP10210K 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.
Sumo2 Conjugation Kit Reference: RP10213K Sumo2 Conjugation Kit is designed for assaying the conjugation of Sumo2 to protein substrates in vitro. In a protein sumoylation reaction, a Sumo protein is first activated by the Sumo activating enzyme SAE1/SAE2 (E1) in an ATP hydrolysis-dependent manner; activated Sumo is then transferred to the Sumo conjugating enzyme Ube2I (E2) by formation of a thiolester bond with the catalytic cysteine residue of Ube2I; Sumo-charged Ube2I can mediate protein sumoylation in the presence of an appropriate Sumo ligase (E3).
Sumo3 Conjugation Kit Reference: RP10214K Sumo3 Conjugation Kit is designed for assaying the conjugation of Sumo3 to protein substrates in vitro. In a protein sumoylation reaction, a Sumo protein is first activated by the Sumo activating enzyme SAE1/SAE2 (E1) in an ATP hydrolysis-dependent manner; activated Sumo is then transferred to the Sumo conjugating enzyme Ube2I (E2) by formation of a thiolester bond with the catalytic cysteine residue of Ube2I; Sumo-charged Ube2I can mediate protein sumoylation in the presence of an appropriate Sumo ligase (E3).
Sumo1, Sumo2, Sumo 3 Conjugation Kit Reference: RP10215K Sumo1, Sumo2, Sumo 3 Conjugation Kit is designed for assaying the conjugation of Sumo1, Sumo2 and Sumo3 to protein substrates in vitro. In a protein sumoylation reaction, a Sumo protein is first activated by the Sumo activating enzyme SAE1/SAE2 (E1) in an ATP hydrolysis-dependent manner; activated Sumo is then transferred to the Sumo conjugating enzyme Ube2I (E2) by formation of a thiolester bond with the catalytic cysteine residue of Ube2I; Sumo-charged Ube2I can mediate protein sumoylation in the presence of an appropriate Sumo ligase (E3).
Ubiquitin Linkage Screening Kit II Reference: RP10218K Ubiquitin Linkage Screening Kit II is designed for two purposes: 1) to identify the ubiquitin lysine residue that is used to link the polyubiquitin chain conjugated on a protein substrate; 2) to determine whether a protein substrate is polyubiquitinated or conjugated with multiple monoubiquitin.
Proteasome Activity Fluorometric Assay Kit I Reference: RP10220K Proteasome Activity Fluorometric Assay Kit I is designed for assaying the proteasome’s chymotrypsin-like activity in vitro using purified proteasome, cleared cell lysates or tissue extracts. The proteasome cleaves SUC-LLVY-AMC and the released AMC fluorescence can be monitored using a plate reader or fluorometer at the excitation/emission wavelength of 360nm/460nm, respectively.
Proteasome Activity Fluorometric Assay Kit II Reference: RP10221K Proteasome Activity Fluorometric Assay Kit II is designed for assaying the proteasome’s chymotrypsin-like, trypsin-like and caspase-like activities in vitro using purified proteasome, cleared cell lysates or tissue extracts. The proteasome cleaves the supplied 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.
Immunoproteasome Activity Fluorometirc Assay Kit I Reference: RP10222K Immunoproteasome Activity Fluorometric Assay Kit I was designed for assaying the immunoproteasomes'' chymotrypsin-like activity in vitro using purified proteasome, cell lysates or tissue extracts. The proteasome cleaves Ac-ANW-AMC and the released AMC fluorescence can be monitored using a plate reader or fluorometer at the excitation/emission wavelength of 360nm/460nm, respectively.
Rapid 26S Proteasome Purification Kit-L Reference: RP10225K The Rapid 26S Proteasome Purification Kit is designed to rapidly purify the endogenous 26S proteasome from cell or tissue extracts within 8 hours according to a published method (reference 1) with modifications. This approach uses the N-terminal ubiquitin-like (Ubl) domain of human RAD23B as an affinity bait, which allows the rapid and gentle isolation of endogenous 26S proteasomes. The bound 26S proteasome is subsequently eluted using the C-terminal two ubiquitin-interacting motifs (UIM) of human S5a
26S Proteasome-mediated Protein Degradation Kit Reference: RP10226K This kit is designed for assaying 26S proteasome-mediated degradation of ubiquitinated proteins in vitro. It contains highly purified active bovine 26S proteasome that has been used by different research groups for assaying protein degradation.
K63 Polyubiquitin Chain Capture Kit Reference: RP10230K This kit is designed for enrichment of polyubiquitinated cellular proteins conjugated with lysine 63 using GST-TAB2 (NZF). Polyubiquitinated proteins bound on GST-TAB2 (NZF) can be precipitated using glutathione resin and then eluted by a buffer containing 10 mM glutathione. Polyubiquitinated proteins can be assayed by immunoblotting or mass spectrometry