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Otubain-1 (human), (recombinant) (His-tag)

 
BML-UW0680-0100 100 µg 258.00 USD
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Product Specification

Alternative Name:OTUB1, Ubiquitin thioesterase OTUB1, Ubiquitin-specific-processing protease OTUB1, Deubiquitinating enzyme OTUB1
 
MW:~33kDa
 
Source:Produced in E. coli.
 
EC:3.1.2.-
 
UniProt ID:Q96FW1
 
Formulation:Liquid. In 50mM TRIS, pH 7.5, containing 50mM NaCl and 1mM DTT.
 
Purity:≥95% (SDS-PAGE)
 
Application Notes:Uses:
1. Deubiquitinylation assays using polyubiquitin chains or ubiquitin-protein conjugates.
2. Studies of the K48-linkage specific polyubiquitin chain binding and deubiquitinylation.
3. Investigation of the role of Otubain-1 in various biological processes.
4. Development of inhibitors for Otubain-1 deubiquitinylating activity.
 
Shipping:Shipped on Dry Ice
 
Short Term Storage:-80°C
 
Long Term Storage:-80°C
 
Use/Stability:Stable for at least 6 months after receipt when stored at -80°C.
 
Handling:Avoid freeze/thaw cycles. After opening, prepare aliquots and store at -80°C.
 
Scientific Background:Otubain-1 belongs to the ovarian tumor (OTU) domain class of cysteine protease deubiquitinating enzymes and has been implicated in mediating lymphocyte antigen responsiveness and may be generally involved in RNA processing and cell adhesion/morphology. Human otubain-1 has been shown to exhibit high linkage-specificity in vitro, cleaving Lys48 (K48)-linked polyubiquitin but not K63-, K29-, K6-, or K11-linked polyubiquitin, or linear α-linked polyubiquitin. Cleavage is not limited to either end of a polyubiquitin chain, and both free and substrate-linked polyubiquitin are disassembled.
 
bml-uw0680 otubain 1 sdspag
Figure 1: SDS-PAGE image of His6-Otubain-1 (Coomassie). Lane 1: MW markers. Lane 2: Otubain-1 (human) (recombinant) (His) (Prod. No. BML-UW0680) (8µg).
bml-uw0680-otubain 1 polyub
Figure 2: Deubiquitinylation assay: Western Blot showing polyubiquitin chains (K48-linked, Prod. No. BML-UW8860; K63-linked, Prod. No. BML-UW9570) incubated with (+) or without (-) Otubain-1 (Prod. No. BML-UW0680), detected with ubiquitin polyclonal antibody (Prod. No. BML-UG9510) (1:1'000)
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bml-uw0680 otubain 1 sdspag bml-uw0680-otubain 1 polyub

General Literature References

Evidence for bidentate substrate binding as the basis for the K48 linkage specificity of otubain 1: T. Wang, et al.; J. Mol. Biol. 386, 1011 (2009), Abstract;
Structural basis and specificity of human otubain 1-mediated deubiquitination: M.J. Edelmann, et al.; Biochem. J. 418, 379 (2009), Abstract;
Two isoforms of otubain 1 regulate T cell anergy via GRAIL: L. Soares, et al.; Nat. Immunol. 5, 45 (2004), Abstract;
Otubains: a new family of cysteine proteases in the ubiquitin pathway: M.Y. Balakirev, et al.; EMBO Rep. 4, 517 (2003), Abstract;

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