The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/10000 - 1/50000. Detects a band of approximately 36 kDa (predicted molecular weight: 36 kDa).
1/10 - 1/100.
1/100 - 1/500. ab172730-Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
Is unsuitable for ICC or IHC-P.
Metalloprotease that specifically cleaves 'Lys-63'-linked polyubiquitin chains (PubMed:19214193, PubMed:20656690, PubMed:24075985, PubMed:26344097). Does not have activity toward 'Lys-48'-linked polyubiquitin chains. Component of the BRCA1-A complex, a complex that specifically recognizes 'Lys-63'-linked ubiquitinated histones H2A and H2AX at DNA lesions sites, leading to target the BRCA1-BARD1 heterodimer to sites of DNA damage at double-strand breaks (DSBs). In the BRCA1-A complex, it specifically removes 'Lys-63'-linked ubiquitin on histones H2A and H2AX, antagonizing the RNF8-dependent ubiquitination at double-strand breaks (DSBs) (PubMed:20656690). Catalytic subunit of the BRISC complex, a multiprotein complex that specifically cleaves 'Lys-63'-linked ubiquitin in various substrates (PubMed:20656690, PubMed:24075985, PubMed:26344097, PubMed:26195665). Mediates the specific 'Lys-63'-specific deubiquitination associated with the COP9 signalosome complex (CSN), via the interaction of the BRISC complex with the CSN complex (PubMed:19214193). The BRISC complex is required for normal mitotic spindle assembly and microtubule attachment to kinetochores via its role in deubiquitinating NUMA1 (PubMed:26195665). Plays a role in interferon signaling via its role in the deubiquitination of the interferon receptor IFNAR1; deubiquitination increases IFNAR1 activity by enhancing its stability and cell surface expression (PubMed:24075985, PubMed:26344097). Down-regulates the response to bacterial lipopolysaccharide (LPS) via its role in IFNAR1 deubiquitination (PubMed:24075985).
Heart, brain, placenta, lung, liver, skeletal muscle, kidney and pancreas. Aberrantly expressed in the vast majority of breast tumors.
Involvement in disease
A chromosomal aberration involving BRCC3 is a cause of pro-lymphocytic T-cell leukemia (T-PLL). Translocation t(X;14)(q28;q11) with TCRA.
Belongs to the peptidase M67A family. BRCC36 subfamily. Contains 1 MPN (JAB/Mov34) domain.
Nucleus. Cytoplasm. Cytoplasm, cytoskeleton, spindle pole. Localizes at sites of DNA damage at double-strand breaks (DSBs) (PubMed:20656690, PubMed:26344097). Interaction with FAM175B/ABRO1 retains BRCC3 in the cytoplasm (PubMed:20656690).
Western blot - Anti-BRCC36 antibody [EPR4366] (ab108411)
Predicted band size : 36 kDa
Lane 1: Wild type HAP1 whole cell lysate (40 µg) Lane 2: Empty Lane Lane 3: BRCC3 knockout HAP1 whole cell lysate (40 µg) Lane 4: MCF7 whole cell lysate (40 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab108411 observed at 36 kDa. Red - loading control, ab18058, observed at 130 kDa.
ab108411 was shown to recognize empty when empty knockout samples were used, along with additional cross-reactive bands. Wild-type and empty knockout samples were subjected to SDS-PAGE. Ab108411 and ab18058 (Mouse anti Vinculin loading control) were incubated overnight at 4°C at 10000 dilution and 1/10000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
Western blot - BRCC36 antibody [EPR4366] (ab108411)
All lanes : Anti-BRCC36 antibody [EPR4366] (ab108411) at 1/10000 dilution
Lane 1 : HeLa cell lysate Lane 2 : MCF7 cell lysate Lane 3 : SKBR-3 cell lysate Lane 4 : 293T cell lysate