Recombinant human Ubiquitin protein Reference: RP10173LQ Ubiquitin (Ub) is a 76 amino acid protein widely expressed in the cytoplasmic and nucleus of cells. Ub is posttranslationally conjugated to proteins by the E1, E2, E3 protein ubiquitination cascade. Ub can be conjugated on proteins as monoUb or polyUb chains. Protein ubiquitination plays both proteolytic and nonproteolytic functions. Usually, polyubiquitinated proteins are targeted to the 26S proteasome for proteolysis. Typical concentration to support in vitro ubiquitination is 50-100 μM.
Recombinant human Ubiquitin(+1) protein Reference: RP10174LQ Ubiquitin (+1) consists of the first 75 amino acids of ubiquitin, but lacks glycine 76 because of the deletion of two of the three nucleotides that encode glycine 76. The resulting frame shift leads to the addition of 20 amino acids (YADLREDPDRQDHHPGSGAQ) after glycine 75. Ubiquitin (+1) can not support ubiquitination reactions. When expressed at high levels, it could inhibit proteasomal degradation. Ubiquitin (+1) was also found in protein aggregates in patient brains of Alzheimer’s disease and other neurodegenerative diseases.
20S proteasome Reference: RP10191TLQ The 20S proteasome has a barrel – shaped structure arranged as four heptomeric rings of αββα. In eukaryotes, each of α and β ring is composed of seven different proteins. The β1, β2 and β5 subunits have ‘caspase-like’, ‘trypsin-like’ and ‘chymotypsin-like’ activities, respectively. In 26S proteasome-mediated protein degradation, to entry the β chamber of the 20S proteasome that houses the proteolytic sites, a substrate protein has to pass through a substrate translocation channel consisting of the double-ring formed by six ATPases of PA700 and the α chamber formed by α subunits of the 20S proteasome.
K6-linked diUb Reference: RP10195D This product is a native K6-linked diUb which can be used as a substrate for deubiquitinating enzymes. It can also be used to investigate linkage specificiy of proteins that contain ubiquitin-associated domains or ubiquitin-interacting motifs. This product is formed by chemical ligation.
N-terminal Biotin-Ubiquitin Reference: RP10199DLQ A single biotin moiety is conjugated on the N-terminus of ubiquitin. All lysine residues of ubiquitin are still available for formation of polyubiquitin chains. The extremely high binding affinity between biotin and streptavidin allows the use of N-terminal biotin-ubiquitin for the following in vitro assays: 1) rapid and efficient purification of polyubiquitinated substrate proteins; 2) monitoring substrate ubiquitination using an HRP-conjugated streptavidin antibody; and 3) setting up high throughput FRET assays to monitor substrate ubiquitination in assays screening E3 ubiquitin ligase inhibitors.
Recombinant human Ubiquitin carboxyl-terminal hydrolase 7/USP7 protein Reference: RP10201LQ USP7 is a cysteine protease, belongs to the family of ubiquitin-specific proteases. USP7 was first discovered as a binding enzyme to the Herpes simplex viral protein. Studies have shown that USP7 could deubiquitinate the autoubiquitination of an E3 ligase called HDM2 that promotes ubiquitination and subsequent degradation of p53. The USP7/HDM2/p53 interaction results in higher protein levels of HDM2 and lower levels of p53. Because of its apparent role in different types of pathologies, including lung and liver cancer, it has become a possible target for drug therapies.
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.