Key features and details
- Rabbit polyclonal to FOXO3A (phospho S253)
- Suitable for: WB, IHC-P, ICC/IF
- Reacts with: Mouse, Human
- Isotype: IgG
Product nameAnti-FOXO3A (phospho S253) antibody
See all FOXO3A primary antibodies
DescriptionRabbit polyclonal to FOXO3A (phospho S253)
SpecificityThis antibody is specific for FOXO3A, only when phosphorylated at serine 253.
Tested applicationsSuitable for: WB, IHC-P, ICC/IFmore details
Species reactivityReacts with: Mouse, Human
Synthetic peptide corresponding to Human FOXO3A.
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Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
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Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Storage bufferpH: 7
Preservative: 0.02% Sodium azide
Constituents: PBS, 50% Glycerol, 0.87% Sodium chloride
Concentration information loading...
PurityImmunogen affinity purified
Purification notesThe antibody was affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific phosphopeptide. The antibody against non-phosphopeptide was removed by chromatography using non-phosphopeptide corresponding to the phosphorylation site.
Our Abpromise guarantee covers the use of ab47285 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/500 - 1/1000. Detects a band of approximately 67, 105 kDa (predicted molecular weight: 71 kDa).|
|IHC-P||Use at an assay dependent concentration.|
|ICC/IF||Use a concentration of 1 µg/ml.|
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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All lanes : Anti-FOXO3A (phospho S253) antibody (ab47285) at 1/500 dilution
Lane 1 : extracts from NIH/3T3 cells untreated
Lane 2 : extracts from NIH/3T3 cells treated with serum
Lysates/proteins at 30 µg per lane.
Predicted band size: 71 kDa
Observed band size: 105,67 kDa why is the actual band size different from the predicted?
1. Add fresh basic medium to the vessels and starve the cells for 18-24 hours.
2. Drain the cultured medium, add growth medium and basic medium containing 10% serum to the vessels.
Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using ab47285 at a 1/50 dilution.
Left image : Untreated
Right image : Treated with P-Peptide
ICC/IF image of ab47285 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab47285, 1µg/ml) overnight at +4°C. 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) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
ab47285 at 1/200 dilution staining FOXO3A in human 293FT cells by Immunocytochemistry/ Immunofluorescence. Cells were fixed in formaldehyde, permebilized in 0.5% Triton X-100 and then blocked in 5% BSA for 1 hour at 25°C. The primary antibody was used at 1/200 dilution in PBS and incubated with sample at 4°C for 12 hours. An Alexa Fluor® 488 conjugated Goat polyclonal to rabbit IgG was used as secondary at 1/500 dilution. Image show green staining with ab47285 and blue with DAPI.
ab47285 has been referenced in 32 publications.
- Lallemand F et al. The high protein expression of FOXO3, but not that of FOXO1, is associated with markers of good prognosis. Sci Rep 10:6920 (2020). PubMed: 32332845
- Lee E et al. Activated pathogenic Th17 lymphocytes induce hypertension following high-fructose intake in Dahl salt-sensitive but not Dahl salt-resistant rats. Dis Model Mech 13:N/A (2020). PubMed: 32179549
- Li J et al. Circular RNA circFBXO11 modulates hepatocellular carcinoma progress and oxaliplatin resistance through miR-605/FOXO3/ABCB1 axis. J Cell Mol Med 24:5152-5161 (2020). PubMed: 32222024
- Xie T et al. microRNA-582 Potentiates Liver and Lung Metastasis of Gastric Carcinoma Cells Through the FOXO3-Mediated PI3K/Akt/Snail Pathway. Cancer Manag Res 12:5201-5212 (2020). PubMed: 32636681
- Lee JW et al. TLR4 (toll-like receptor 4) activation suppresses autophagy through inhibition of FOXO3 and impairs phagocytic capacity of microglia. Autophagy 15:753-770 (2019). PubMed: 30523761