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
Linear Polyubiquitin Chain Capture Kit Reference: RP10231K This kit is designed for enrichment of cellular proteins conjugated with linear polyubiquitin chains using GST-NEMO (UBAN). Polyubiquitinated proteins bound on GST-NEMO (UBAN) can be precipitated using glutathione resin and eluted by a buffer containing 10 mM glutathione. Polyubiquitinated proteins can be assayed by immunoblotting or mass spectrometry.
Polyubiquitin Chain Capture Kit Reference: RP10232K This kit is designed for enrichment of polyubiquitinated cellular proteins using GST tagged recombinant proteins that bind lysine 48 and lysine 63 polyubiquitin chains. It can be used to enrich polyubiquitinated proteins in whole cell or tissue lysates. Bound proteins can be precipitated using glutathione resin and eluted by a buffer containing 10 mM glutathione, and then assayed by im-munoblotting or mass spectrometry.
Deubiquitinating Enzyme (DUB) Identification Kit Reference: RP10237K This kit is designed to enrich deubiquitinating enzymes in crude or partially purified cell/tissue extracts, followed by identification using immunoblotting or mass spectrometry.
Caspase-3, 7 Activity Fluorometric Assay Kit Reference: RP10240K Caspase-3, 7 Activity Fluorometric Assay Kit can be used for assaying caspase-3, 7 activities in cell/tissue extracts in a 96-well plate format. Each of the supplied fluorogenic substrate and the pan caspase inhibitor Z-VAD-FMK is sufficient for 250 x 100 μl reactions. The supplied TRAIL-treated cell lysate is used as a positive control, sufficient for 5 X 100 μl reactions.