1/100 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Component of the eukaryotic translation initiation factor 3 (eIF-3) complex, which is required for several steps in the initiation of protein synthesis. The eIF-3 complex associates with the 40S ribosome and facilitates the recruitment of eIF-1, eIF-1A, eIF-2:GTP:methionyl-tRNAi and eIF-5 to form the 43S pre-initiation complex (43S PIC). The eIF-3 complex stimulates mRNA recruitment to the 43S PIC and scanning of the mRNA for AUG recognition. The eIF-3 complex is also required for disassembly and recycling of post-termination ribosomal complexes and subsequently prevents premature joining of the 40S and 60S ribosomal subunits prior to initiation. Essential for the initiation of translation on type-1 viral ribosomal entry sites (IRESs), like for HCV, PV, EV71 or BEV translation (PubMed:23766293, PubMed:24357634). In case of FCV infection, plays a role in the ribosomal termination-reinitiation event leading to the translation of VP2 (PubMed:18056426).
Belongs to the eIF-3 subunit A family. Contains 1 PCI domain.
Phosphorylated. Phosphorylation is enhanced upon serum stimulation.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human prostate carcinoma (left) and mouse teratoma (right) tissues labelling eIF3A with ab86146 at 1/200 (1µg/ml). Detection: DAB.
Western blot - Anti-eIF3A antibody (ab86146)This image is courtesy of an anonymous Abreview
Anti-eIF3A antibody (ab86146) at 1/1000 dilution + Mouse pancreatic cancer whole cell lysate at 20 µg
Secondary IRDye® 680LT-conjugated donkey anti-mouse IgG polyclonal at 1/15000 dilution
All lanes : Anti-eIF3A antibody (ab86146) 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
Developed using the ECL technique.
Predicted band size: 167 kDa
Exposure time: 3 seconds
Immunoprecipitation - eIF3A antibody (ab86146)
Detection of Human eIF3A by Immunoprecipitation, using ab86146 at 3µg/mg lysate. Image shows immunoprecipitated eIF3A detected with post IP WB, loading 20% of IP and using HeLa whole cell lysate at 1mg, with an antibody recognising an upstream epitope in lane 1, ab86146 at 1 µg/ml in lane 2 and control IgG in lane 3. Detection: Chemiluminescence with an exposure time of 3 seconds.
Conrad KD et al. MicroRNA-122 dependent binding of Ago2 protein to hepatitis C virus RNA is associated with enhanced RNA stability and translation stimulation. PLoS One8:e56272 (2013).
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