1/500 - 1/2000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human prostate carcinoma tissue labelling BRCC36 with ab115172 at 1/1000 (1µg/ml). Detection: DAB.
Western blot - BRCC36 antibody (ab115172)
All lanes : Anti-BRCC36 antibody (ab115172) at 0.1 µg/ml
Lane 1 : HeLa cell lysate at 50 µg Lane 2 : HeLa cell lysate at 15 µg Lane 3 : HeLa cell lysate at 5 µg Lane 4 : 293T cell lysate at 50 µg Lane 5 : Mouse NIH3T3 cell lysate at 50 µg
Predicted band size : 36 kDa
Exposure time : 30 seconds
Immunoprecipitation - BRCC36 antibody (ab115172)
Immunoprecipitation of BRCC36 (3 µg of antibody per mg of HeLa cell lysate; 20% of the IP loaded). Lane 1-Immunoprecipitation with an anti-BRCC36 antibody that recognizes an upstream epitope; Lane 2-Immunoprecipitation with anti-BRCC36 antibody (ab115172); Lane 3-Immunoprecipitation with IgG control. Anti-BRCC36 antibody (ab115172) used at 0.4 µg/ml for western blot. Detection: Chemiluminescence with exposure time of 30 sec.
Jamsai D et al. Loss of GGN leads to pre-implantation embryonic lethality and compromised male meiotic DNA double strand break repair in the mouse. PLoS One8:e56955 (2013).
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