Anti-FOXO3A (phospho S253) antibody (ab47285)
- Datasheet
- References (16)
- Protocols
Overview
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Product nameAnti-FOXO3A (phospho S253) antibody
See all FOXO3A primary antibodies -
DescriptionRabbit polyclonal to FOXO3A (phospho S253)
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Host speciesRabbit
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SpecificityThis antibody is specific for FOXO3A, only when phosphorylated at serine 253.
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Tested applicationsSuitable for: WB, IHC-Fr, IHC-P, ELISA, ICC/IFmore details
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Species reactivityReacts with: Mouse, Rat, Human
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Immunogen
Synthetic peptide corresponding to Human FOXO3A.
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Positive control
- Human breast carcinoma tissue; NIH/3T3 cell extracts
Properties
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FormLiquid
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Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
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Storage bufferpH: 7.40
Preservative: 0.02% Sodium azide
Constituents: PBS, 50% Glycerol, 0.87% Sodium chloride -
Concentration information loading... -
PurityImmunogen affinity purified
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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.
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ClonalityPolyclonal
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IsotypeIgG
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Research areas
Associated products
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Compatible Secondaries
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Positive Controls
Applications
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.
| Application | Abreviews | Notes |
|---|---|---|
| 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. | |
| ELISA | 1/20000. | |
| ICC/IF | Use a concentration of 1 µg/ml. |
Target
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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'.
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Tissue specificityUbiquitous.
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Involvement in diseaseNote=A chromosomal aberration involving FOXO3 is found in secondary acute leukemias. Translocation t(6;11)(q21;q23) with MLL/HRX.
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Sequence similaritiesContains 1 fork-head DNA-binding domain.
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Post-translational
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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Database links
- Entrez Gene: 2309 Human
- Entrez Gene: 56484 Mouse
- Entrez Gene: 294515 Rat
- Omim: 602681 Human
- SwissProt: O43524 Human
- SwissProt: Q9WVH4 Mouse
- Unigene: 220950 Human
- Unigene: 338613 Mouse
see all -
Alternative names
- AF6q21 antibody
- AF6q21 protein antibody
- DKFZp781A0677 antibody
see all
Images
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Western blot - Anti-FOXO3A (phospho S253) antibody (ab47285)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?
Serum treatment:-
1. Add fresh basic medium to the vessels and starve the cells for 18-24 hours.
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2. Drain the cultured medium, add growth medium and basic medium containing 10% serum to the vessels.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXO3A (phospho S253) antibody (ab47285)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
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Immunocytochemistry/ Immunofluorescence - Anti-FOXO3A (phospho S253) antibody (ab47285)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. -
Immunocytochemistry/ Immunofluorescence - Anti-FOXO3A (phospho S253) antibody (ab47285)This image is courtesy of an anonymous Abreview.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.
Protocols
References
This product has been referenced in:
- Zhang H et al. The mechanisms involved in miR-9 regulated apoptosis in cervical cancer by targeting FOXO3. Biomed Pharmacother 102:626-632 (2018). Read more (PubMed: 29602130) »
- Liu S et al. FoxO3a plays a key role in the protective effects of pomegranate peel extract against amikacin-induced ototoxicity. Int J Mol Med 40:175-181 (2017). WB, IHC . Read more (PubMed: 28560451) »
