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 80 kDa (predicted molecular weight: 80 kDa).
1/250 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. Heat up to 98 degrees C, below boiling, and then let cool for 10-20 min.
1/50 - 1/100.
Application notesIs unsuitable for Flow Cyt or IP.
FunctionUbiquitin-binding protein that specifically recognizes and binds 'Lys-63'-linked ubiquitin. Plays a central role in the BRCA1-A complex by specifically binding '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). The BRCA1-A complex also possesses deubiquitinase activity that specifically removes 'Lys-63'-linked ubiquitin on histones H2A and H2AX. Also weakly binds monoubiquitin but with much less affinity than 'Lys-63'-linked ubiquitin. May interact with monoubiquitinated histones H2A and H2B; the relevance of such results is however unclear in vivo. Does not bind Lys-48'-linked ubiquitin. May indirectly Act as a transcriptional repressor by inhibiting the interaction of NR6A1 with the corepressor NCOR1.
Tissue specificityExpressed in testis, ovary, thymus and heart. Expressed in germ cells of the testis.
Sequence similaritiesBelongs to the RAP80 family. Contains 2 UIM (ubiquitin-interacting motif) repeats.
DomainThe UIM-linker region between the 2 UIM repeats determines the selectivity for 'Lys-63'-linked ubiquitin. The length of the linker is important. The linker reduces the flexibility between the UIM repeats and promotes high-affinity and linkage-selective interactions. The Abraxas-interacting region (AIR) mediates the interaction with FAM175A/Abraxas.
Post-translational modificationsSumoylated. Phosphorylated upon DNA damage by ATM or ATR.
Cellular localizationNucleus. Localizes at sites of DNA damage at double-strand breaks.
ab124763, at 1/250, staining RAP80 in paraffin-embedded Human testis tissue by Immunohistochemistry.
References for Anti-RAP80 antibody [EPR5315] (ab124763)
This product has been referenced in:
Hu B et al. HSCARG, a novel regulator of H2A ubiquitination by downregulating PRC1 ubiquitin E3 ligase activity, is essential for cell proliferation. Nucleic Acids Res42:5582-93 (2014).
Read more (PubMed: 24711370) »
Kakarougkas A et al. Co-operation of BRCA1 and POH1 relieves the barriers posed by 53BP1 and RAP80 to resection. Nucleic Acids ResN/A:N/A (2013).
Read more (PubMed: 24013561) »