1/100 - 1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
1/50 - 1/500.
Tyrosine phosphatase that specifically dephosphorylates 'Tyr-142' of histone H2AX (H2AXY142ph). 'Tyr-142' phosphorylation of histone H2AX plays a central role in DNA repair and acts as a mark that distinguishes between apoptotic and repair responses to genotoxic stress. Promotes efficient DNA repair by dephosphorylating H2AX, promoting the recruitment of DNA repair complexes containing MDC1. Its function as histone phosphatase probably explains its role in transcription regulation during organogenesis. Coactivates SIX1, and seems to coactivate SIX2, SIX4 and SIX5. The repression of precursor cell proliferation in myoblasts by SIX1 is switched to activation through recruitment of EYA3 to the SIX1-DACH1 complex and seems to be dependent on EYA3 phosphatase activity (By similarity). May be involved in development of the eye.
Belongs to the HAD-like hydrolase superfamily. EYA family.
Phosphorylated upon DNA damage, probably by ATM or ATR. Ser-266 phosphorylation is required for localization at sites of DNA damage and directing interaction with H2AX.
Cytoplasm. Nucleus. Localizes at sites of DNA damage at double-strand breaks (DSBs). With decreasing efficiency, translocalized to the nucleus by SIX2 and SIX5, and SIX4, respectively.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma tissue labelling EYA3 with ab95876 at 1/200 (1µg/ml). Detection: DAB.
Western blot - EYA3 antibody (ab95876)
All lanes : Anti-EYA3 antibody (ab95876) at 0.04 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
Developed using the ECL technique
Predicted band size : 63 kDa
Exposure time : 30 seconds
Immunoprecipitation - EYA3 antibody (ab95876)
Detection of human Eya3 by Western Blot of Immunoprecipitate.
ab95876 at 0.04µg/ml staining Eya3 in HeLa cell lysate immunoprecipitated using ab95876 at 3µg/mg (1mg/IP; 20% of IP loaded/lane).
Detection: Chemiluminescence with exposure time of 10 seconds.