Biotin-Ub-PA Reference: RP10243D Biotin-Ub-PA is a newly developed potent and specific inhibitor of deubiquitinating enzymes (DUBs), which has a N-terminal biotin tag. An aminohexanoic acid (Ahx) linker is used to create extra space between the biotin and Ub protein for efficient access of the biotin binding entity. This activity probe can be used for activity profiling experiments and determining DUB inhibitor specificity. It has two unique capabilities: it forms a covalent linkage with the active site Cys residue of a DUB that can be cleaved by acid treatment (5% aq. TFA), allowing for proteomic analyses. Biotin-Ub-PA targets all three major DUB families: UCH, USP and OTU
Cy5-Ub-VME Reference: RP10245D Cy5-Ub-VME is a potent, irreversible and specific inhibitor of deubiquitinating enzymes (DUBs), which is labeled on the N-terminus with a Cy5 dye (Cy5, Ex 625-650 nm, Em 670 nm). This ubiquitin-based activity probe can be used for activity profiling experiments and labeling of DUBs with a fluorescent dye (Cy5). Cy5 labelling allows for detection of DUB labeling by in-gel fluorescence.
biotin-SUMO2-VME Reference: RP10246D A potent, irreversible and specific inhibitor of deSUMOylation proteases (SENPs) that cleave SUMO2 conjugates. It is N-terminally tagged with a biotin, which allows for sensitive identification of SENPs using HRP-conjugated streptavidin in immunoblotting assays, or purification of SENPs using streptavidin resin. The biotin tag is separated from SUMO2 by an aminohexanoic acid (Ahx) linker that could increase the reactivity of the biotin moiety. In addition, Cys48 in SUMO2 was mutated to Ser.
Ub-Rhodamine110 Reference: RP10248DLQ Ub-Rhodamine 110 is a quenched, fluorescent substrate for deubiquitinating enzymes. Cleavage of the amide bond between the C-terminal glycine of ubiquitin and rhodamine110 results in an increase in rhodamine fluorescence at 535 nm (Exc. 485 nm).
Recombinant human Ubiquitin thioesterase ZRANB1 protein Reference: RP10005LQ TRABID is an OUT domain-containing deubiquitinating enzyme that exhibits specificity to K29/K33-linked polyubiquitin chains. 6xHis-GST-TRABID(NZF1) contains the N-terminal NZF1 domain (amino acid 1-33) of TRABID fused at the C-terminal of the 6xHis-GST tag. TRABID(NZF1) specifically binds K29/K33 polyubiquitin chains. This protein can be used to pull down K29 and K33 polyubiquitin chains using Ni-NTA resin or glutathione resin.
Recombinant human Proteasome inhibitor PI31 subunit protein Reference: RP10021LQ PI31 is a proline-rich protein and its C-terminal region can act as a potent 20S proteasome inhibitor in vitro. In cells, it plays a role in regulating 26S proteasome assembly and activity.
Recombinant human SENP1 (catalytic domain) protein Reference: RP10024LQ SUMO1-3 (small ubiquitin-related modifiers) are ubiquitin-like proteins that can be conjugated to substrate proteins using a cascade of enzymatic reactions. Protein sumoylation can be reversed by the sentrin-specific (SUMO is also named sentrin) proteases (SENPs). In human cells, SENPs have six members including SENP1, SENP2, SENP3, SENP5, SENP6 and SENP7. SENPs mostly reside in the nucleus. They function to generate mature SUMO1-3 from the SUMO precursors and attenuate signals mediated by sumoylation. The SENP1 catalytic domain comprises amino acid from 419 to 644.
Recombinant human Ubiquitin-conjugating enzyme E2 S/UBE2S(1-156) protein Reference: RP10058LQ UbE2S is an E2 enzyme, which is part of the E1, E2, and E3 cascade responsible for ubiquitination of protein substrates. UbE2S works with the Ub ligase, anaphase-promoting complex (APC/C), to ubiquitinate proteins like Cyclin B1. UbE2S was found to mediate synthesis of K11-specific polyubiquitin chains. UbE2S(1-156) lacks the C-terminal region of UbE2S, and this truncated version is more efficient than UbE2S in catalyzing synthesis of K11-linked polyubiquitin chains.
Recombinant human Ubiquitin (K48R) protein Reference: RP10077LQ 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. In Ub(K48R), lysine 48 was substituted with an arginine, thereby it cannot form lysine 48-linked polyubiquitin chains.
Recombinant human Ubiquitin carboxyl-terminal hydrolase 14 protein Reference: RP10085LQ Usp14 is one of the deubiquitinating enzymes of the human 26S proteasome. Usp14 belongs to the Ub-specific proteases family. Usp14 catalyzes Ub chain trimming that shortens a polyUb chain from the distal end of the Ub chain. Usp14 was found to play an essential role in maintaining the protein levels of monomeric Ub in cells. Interestingly, Usp14 was also found to inhibit proteasomal degradation independent of its deubiquitinating activity.
Recombinant human ADRM1&UCH-L5 protein Reference: RP10087LQ Adrm1 is an integral Ub receptor of the 26S proteasome. Adrm1 binds both Ub and polyUb chains via its N-terminal Pru domain. The Pru domain also binds S1 to locate Adrm1 on the 26S proteasome. The C-terminal domain of Adrm1 binds and activates the deubiquitinating enzyme Uch37, though which Uch37 is recruited to the 26S proteasome. Currently, it is not known whether Adrm1 activates Uch37 to deubiquitinate polyUb chains.