The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/200 - 1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Use a concentration of 1 - 4 µg/ml.
Recommend PFA Fixation and Triton X-100 treatment
FunctionHydrolase that can remove conjugated ubiquitin from proteins and participates in endoplasmic reticulum-associated degradation (ERAD) for misfolded lumenal proteins. May act by triming the ubiquitin chain on the associated substrate to facilitate their threading through the VCP/p97 pore. Ubiquitin moieties on substrates may present a steric impediment to the threading process when the substrate is transferred to the VCP pore and threaded through VCP's axial channel. Mediates deubiquitination of both 'Lys-48'- and 'Lys-63'-linked polyubiquitin chains. Able to cleave both polyubiquitin and di-ubiquitin.
Sequence similaritiesContains 1 C2H2-type zinc finger. Contains 1 OTU domain.
DomainThe UBAX-like region mediates the interaction with VCP. According to PubMed:19818707, it corresponds to a UBX domain, which is a hallmark for VCP-associated proteins. However, no canonical UBX is detected by prediction tools such as Pfam or PROSITE.
Immunofluorescent staining of Human cell line U-2 OS shows positivity in nucleus but not nucleoli, plasma membrane and cytoskeleton (actin filaments). Recommended concentration of ab121984 1-4 µg/ml. Cells treated with PFA/Triton X-100.