1/100 - 1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Plays a critical role in both constitutive and enhancer-dependent splicing by mediating protein-protein interactions and protein-RNA interactions required for accurate 3'-splice site selection. Recruits U2 snRNP to the branch point. Directly mediates interactions between U2AF2 and proteins bound to the enhancers and thus may function as a bridge between U2AF2 and the enhancer complex to recruit it to the adjacent intron.
Belongs to the splicing factor SR family. Contains 2 C3H1-type zinc fingers. Contains 1 RRM (RNA recognition motif) domain.
The C-terminal SR-rich domain is required for interactions with SR proteins and the splicing regulators TRA and TRA2, and the N-terminal domain is required for formation of the U2AF1/U2AF2 heterodimer.
Phosphorylated upon DNA damage, probably by ATM or ATR.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human lung carcinoma (left) and mouse CT26 colon carcinoma (right) tissues labelling U2AF35 with ab86305 at 1/200 (1µg/ml). Detection: DAB.
Western blot - Anti-U2AF35 antibody (ab86305)
All lanes : Anti-U2AF35 antibody (ab86305) at 0.04 µg/ml
Lane 1 : HeLa whole cell lysate at 50 µg Lane 2 : HeLa whole cell lysate at 15 µg Lane 3 : HeLa whole cell lysate at 5 µg Lane 4 : 293T whole cell lysate at 50 µg Lane 5 : NIH3T3 whole cell lysate at 50 µg
Detection of U2AF35 by Western Blot of Immunprecipitate.
ab86305, at 1 µg/ml, staining U2AF35 in HeLa whole cell lysate immunoprecipitated using ab86305 at 3 µg/mg lysate (1 mg/IP; 20% of IP loaded/lane). Detection: Chemiluminescence with an exposure time of 10 seconds.
Martinez NM et al. Widespread JNK-dependent alternative splicing induces a positive feedback loop through CELF2-mediated regulation of MKK7 during T-cell activation. Genes Dev29:2054-66 (2015).
Read more (PubMed: 26443849) »
Zhang J et al. Disease-associated mutation in SRSF2 misregulates splicing by altering RNA-binding affinities. Proc Natl Acad Sci U S A112:E4726-34 (2015).
Read more (PubMed: 26261309) »