1/100 - 1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Component the THO subcomplex of the TREX complex. The TREX complex specifically associates with spliced mRNA and not with unspliced pre-mRNA. It is recruited to spliced mRNAs by a transcription-independent mechanism. Binds to mRNA upstream of the exon-junction complex (EJC) and is recruited in a splicing- and cap-dependent manner to a region near the 5' end of the mRNA where it functions in mRNA export. The recruitment occurs via an interaction between ALYREF/THOC4 and the cap-binding protein NCBP1. The TREX complex is essential for the export of Kaposi's sarcoma-associated herpesvirus (KSHV) intronless mRNAs and infectious virus production. The recruitment of the TREX complex to the intronless viral mRNA occurs via an interaction beteen KSHV ORF57 protein and ALYREF/THOC4. DDX39B functions as a bridge between ALYREF/THOC4 and the THO complex. THOC5 in conjunction with ALYREF/THOC4 functions in NXF1-NXT1 mediated nuclear export of HSP70 mRNA. Regulates the expression of myeloid transcription factors CEBPA, CEBPB and GAB2 by enhancing the levels of phosphatidylinositol 3,4,5-trisphosphate. May be involved in the differentiation of granulocytes and adipocytes. Essential for hematopoietic primitive cell survival and plays an integral role in monocytic development.
Belongs to the THOC5 family.
Phosphorylated on tyrosine upon binding to activated CSF1R; which causes a dissociation of the two proteins. Phosphorylation on Ser-5 and/or Ser-6 is required for nuclear export. Phosphorylated on Thr-328 in insulin-stimulated adipocytes (By similarity). Phosphorylated upon DNA damage, probably by ATM or ATR.
Nucleus. Cytoplasm. Shuttles between nucleus and cytoplasm.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human ovarian carcinoma tissue labelling THOC5 with ab86074 at 1/200 (1µg/ml). Detection: DAB.
Western blot - THOC5 antibody (ab86074)
All lanes : Anti-THOC5 antibody (ab86074) 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 cell lysate at 50 µg Lane 5 : mouse NIH3T3 cell lysate at 50 µg
Developed using the ECL technique.
Predicted band size: 79 kDa Observed band size: 79 kDa
Exposure time: 30 seconds
Immunoprecipitation - THOC5 antibody (ab86074)
Immunoprecipitation/ Western Blot of ab86074:
Whole cell lysate from Hela cells at 1mg for IP, 20% of IP loaded and ab86074 used for IP at 3 µg/mg lysate and 0.4µg/ml for WB. Chemiluminescence with an exposure time of 10 seconds.