Product nameAnti-FOXO3A (phospho S253) antibody
See all FOXO3A primary antibodies
DescriptionRabbit polyclonal to FOXO3A (phospho S253)
Tested applicationsSuitable for: WB, ICC/IF, IHC-P, IPmore details
Species reactivityReacts with: Mouse, Rat
Predicted to work with: Human, Pig, Zebrafish
Synthetic peptide corresponding to Human FOXO3A aa 200-300 conjugated to keyhole limpet haemocyanin.
(Peptide available as
- Recombinant Human FOXO3A protein (ab114191) can be used as a positive control in WB. This antibody gave a positive signal in PC12 (Rat adrenal pheochromocytoma cell line) Cytoplasmic Lysate.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
Storage bufferpH: 7.40
Preservative: 0.02% Sodium azide
Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
Concentration information loading...
PurityImmunogen affinity purified
Immunizing Peptide (Blocking)
Our Abpromise guarantee covers the use of ab31109 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
IHC-P: Use at an assay dependent dilution (PMID 20140201).
WB: Use at a concentration of 1 µg/ml. Detects a band of approximately 63 kDa (predicted molecular weight: 70 kDa). Can be blocked with FOXO3A (phospho S253) peptide (ab27885).
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
FunctionTranscriptional 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'.
Involvement in diseaseNote=A chromosomal aberration involving FOXO3 is found in secondary acute leukemias. Translocation t(6;11)(q21;q23) with MLL/HRX.
Sequence similaritiesContains 1 fork-head DNA-binding domain.
modificationsIn 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.
Cellular localizationCytoplasm > cytosol. Nucleus. Translocates to the nucleus upon oxidative stress and in the absence of survival factors.
- Information by UniProt
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FOXO3A was immunoprecipitated using 0.5mg PC12 whole cell extract, 5µg of Rabbit polyclonal to FOXO3A and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, PC12 whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70°C; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab31109.
Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
Band: 63kDa; FOXO3A
ICC/IF image of ab31109 stained PC12 cells. The cells were 4% PFA fixed (10 min), permabilised in 0.1% PBS-Tween (20 min) and incubated with the antibody (ab31109, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).
ab31109 staining FOXO3A in adenomas from ApcMin/+ mice (30 weeks old) by Immunohistochemistry (paraffin embedded sections). Intestines were opened longitudinally, cleaned, Swiss-rolled, fixed with periodate-lysine-paraformaldehyde, and embedded in paraffin.
Phosphorylated FOXO3A was identified utilizing ab31109, followed by HRP-conjugated goat-anti-rabbit IgG. All stains were then developed in 3, 3′-diaminobenzidine and a haematoxylin counterstain was applied.
This product has been referenced in:
- Zeng X et al. Acylated and unacylated ghrelin inhibit atrophy in myotubes co-cultured with colon carcinoma cells. Oncotarget 8:72872-72885 (2017). Read more (PubMed: 29069832) »
- Gravina GL et al. Pharmacological treatment with inhibitors of nuclear export enhances the antitumor activity of docetaxel in human prostate cancer. Oncotarget 8:111225-111245 (2017). Read more (PubMed: 29340049) »