- Datasheet
- Specific References (3)
- 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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Tested applicationsSuitable for: WB, ICC/IF, IHC-P, IPmore details
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Species reactivityReacts with: Mouse, Rat
Predicted to work with: Human, Pig, Zebrafish
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Immunogen
Synthetic peptide conjugated to KLH derived from within residues 200 - 300 of Human FOXO3A.
(Peptide available as ab27885.)
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Positive control
- This antibody gave a positive signal in PC12 (Rat adrenal pheochromocytoma cell line) Cytoplasmic Lysate.
Properties
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FormLiquid
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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.
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Storage bufferPreservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4 -
Concentration information loading... -
PurityImmunogen affinity purified
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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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Control Peptide
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Immunizing Peptide (Blocking)
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Positive Controls
Applications
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.
| Application | Abreviews | Notes |
|---|---|---|
| WB | ||
| ICC/IF | ||
| IHC-P | ||
| IP |
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.
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: 733621 Pig
- Entrez Gene: 294515 Rat
- Omim: 602681 Human
- SwissProt: O43524 Human
- SwissProt: Q9WVH4 Mouse
- Unigene: 220950 Human
see all -
Alternative names
- AF6q21 antibody
- AF6q21 protein antibody
- DKFZp781A0677 antibody
see all
Anti-FOXO3A (phospho S253) antibody images
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Immunoprecipitation - Anti-FOXO3A (phospho S253) antibody (ab31109)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
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Western blot - FOXO3A (phospho S253) antibody (ab31109)
Performed under reducing conditions.
Predicted band size : 70 kDa -
Immunocytochemistry/ Immunofluorescence - FOXO3A (phospho S253) antibody (ab31109)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). -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - FOXO3A (phospho S253) antibody (ab31109)Image from Guillen-Ahlers H et al, PLoS One. 2010 Feb 5;5(2):e9070, Fig 6.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.
Protocols
References for Anti-FOXO3A (phospho S253) antibody (ab31109)
This product has been referenced in:
- Tenland E et al. Innate Immune Responses after Airway Epithelial Stimulation with Mycobacterium bovis Bacille-Calmette Guérin. PLoS One 11:e0164431 (2016). WB ; Human . Read more (PubMed: 27723804) »
- Guillen-Ahlers H et al. Fas/CD95 Deficiency in Apc Mice Increases Intestinal Tumor Burden. PLoS One 5:e9070 (2010). IHC-P ; Mouse . Read more (PubMed: 20140201) »
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"