The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
IP: Use at 2-5µg/mg of lysate.
WB: 1/1000 - 1/10000. Detects a band of approximately 100 kDa (predicted molecular weight: 80 kDa).
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
Ubiquitin-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.
Expressed in testis, ovary, thymus and heart. Expressed in germ cells of the testis.
Belongs to the RAP80 family. Contains 2 UIM (ubiquitin-interacting motif) repeats.
The 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.
Sumoylated. Phosphorylated upon DNA damage by ATM or ATR.
Nucleus. Localizes at sites of DNA damage at double-strand breaks.
Detection of Human RAP80 by Immunoprecipitation in Whole cell lysate from HeLa cells (1 mg for IP, 20% of IP loaded), using ab70822 at 3 µg/mg lysate for IP and at 1 µg/ml for subsequent Western blot detection.