Key features and details
- Rabbit polyclonal to FOXO3A (phospho S253)
- Suitable for: WB, IHC-Fr, IHC-P, ELISA, ICC/IF
- Reacts with: Mouse, Rat, 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-Fr, IHC-P, ELISA, ICC/IFmore details
Species reactivityReacts with: Mouse, Rat, Human
Synthetic peptide corresponding to Human FOXO3A.
- Human breast carcinoma tissue; NIH/3T3 cell extracts
Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Storage bufferpH: 7.40
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-Fr||Use at an assay dependent concentration. See Abreview.|
|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
- AF6q21 antibody
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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 28 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
- Shan Z et al. CTCF regulates the FoxO signaling pathway to affect the progression of prostate cancer. J Cell Mol Med 23:3130-3139 (2019). PubMed: 30873749
- Shang L et al. MicroRNA-148a-3p promotes survival and migration of endothelial cells isolated from Apoe deficient mice through restricting circular RNA 0003575. Gene 711:143948 (2019). PubMed: 31255737
- Zhou LL et al. Differentially expressed long noncoding RNAs and regulatory mechanism of LINC02407 in human gastric adenocarcinoma. World J Gastroenterol 25:5973-5990 (2019). PubMed: 31660034
- Liang C et al. LncRNA GAS6-AS2 facilitates tumor growth and metastasis of hepatocellular carcinoma by activating the PI3K/AKT/FoxO3a signaling pathway. Int J Clin Exp Pathol 12:4011-4023 (2019). PubMed: 31933797