The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/5000 - 1/15000. Detects a band of approximately 85 kDa (predicted molecular weight: 71 kDa).
Use at 2-5 µg/mg of lysate.
1/200 - 1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Transcriptional activator which triggers apoptosis in the absence of survival factors, including neuronal cell death upon oxidative stress. Recognizes and binds to the DNA sequence 5'-[AG]TAAA[TC]A-3'.
Note=A chromosomal aberration involving FOXO3 is found in secondary acute leukemias. Translocation t(6;11)(q21;q23) with MLL/HRX.
Contains 1 fork-head DNA-binding domain.
In the presence of survival factors such as IGF-1, phosphorylated on Thr-32 and Ser-253 by AKT1/PKB. This phosphorylated form then interacts with 14-3-3 proteins and is retained in the cytoplasm. Survival factor withdrawal induces dephosphorylation and promotes translocation to the nucleus where the dephosphorylated protein induces transcription of target genes and triggers apoptosis. Although AKT1/PKB doesn't appear to phosphorylate Ser-315 directly, it may activate other kinases that trigger phosphorylation at this residue. Phosphorylated by STK4 on Ser-209 upon oxidative stress, which leads to dissociation from YWHAB/14-3-3-beta and nuclear translocation. Phosphorylated by PIM1.
Cytoplasm > cytosol. Nucleus. Translocates to the nucleus upon oxidative stress and in the absence of survival factors.
Lane 1 : MCF7 Cell Lysate Immunoprecipitated with ab70314 at 6ug/mg.
Lane 2: MCF7 Cell Lysate Immunoprecipitated with ab70315 6ug/mg
Lane 3:MCF7 Cell Lysate Immunoprecipitated with Control IgG at 6ug/mg
Western Blot carried out using ab70314 at 1ug/ml
Decction: ECL for 3 minutes.
All lanes : Anti-FOXO3A antibody (ab70314) at 1 µg/ml
Lane 1 : MCF7 Cell Lysate Immunoprecipitated with ab70314 at 6ug/mg Lane 2 : MCF7 Cell Lysate Immunoprecipitated with ab70315 6ug/mg Lane 3 : MCF7 Cell Lysate Immunoprecipitated with Control IgG at 6ug/mg
Developed using the ECL technique.
Exposure time: 3 minutes
Western blot - Anti-FOXO3A antibody (ab70314)
All lanes : Anti-FOXO3A antibody (ab70314) at 0.4 µg/ml
Lane 1 : 293T cell Lysate Lane 2 : Jurkat cell Lysate Lane 3 : Mouse TCMK cell Lysate
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human ovarian carcinoma (left) and mouse renal cell carcinoma (right) tissues labelling FOXO3A with ab70314 at 1/500 (2µg/ml). Detection: DAB.