- Datasheet
- Specific References
- Protocols
Overview
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Product nameAnti-FOXO3A (phospho T32) antibody
See all FOXO3A primary antibodies -
DescriptionRabbit polyclonal to FOXO3A (phospho T32)
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Host speciesRabbit
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Tested applicationsSuitable for: WB, IHC-Pmore details
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Species reactivityReacts with: Human
Predicted to work with: Mouse, Chicken, Xenopus laevis, Zebrafish
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Immunogen
Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human FOXO3A, phosphorylated at T32.
(Peptide available as ab27877.)
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Positive control
- This antibody gave a positive signal in the following whole cell lysates; HT29; Jurkat; 293T Cell extract - treated with Calyculin A,
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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Immunizing Peptide (Blocking)
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Positive Controls
Applications
Our Abpromise guarantee covers the use of ab26649 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 | Use a concentration of 1 µg/ml. Detects a band of approximately 80 kDa (predicted molecular weight: 89,90 kDa). | |
| IHC-P | Use at an assay dependent concentration. see image below |
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: 444992 Xenopus laevis
- Omim: 602681 Human
- SwissProt: O43524 Human
- SwissProt: Q9WVH4 Mouse
- SwissProt: Q6EUW1 Xenopus laevis
- Unigene: 220950 Human
see all -
Alternative names
- AF6q21 antibody
- AF6q21 protein antibody
- DKFZp781A0677 antibody
see all
Images
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Western blot - Anti-FOXO3A (phospho T32) antibody (ab26649)All lanes : Anti-FOXO3A (phospho T32) antibody (ab26649) at 1 µg/ml
Lane 1 :HT29 whole cell lysate (ab3952)
Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
Lane 3 : 293T Cell extract - treated with Calyculin A
Lane 4 :HT29 whole cell lysate (ab3952) with Human FOXO3A (phospho T32) peptide (ab27877) at 1 mg/ml
Lane 5 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate withHuman FOXO3A (phospho T32) peptide (ab27877) at 1 mg/ml
Lane 6 : 293T Cell extract - treated with Calyculin A withHuman FOXO3A (phospho T32) peptide (ab27877) at 1 mg/ml
Lane 7 :HT29 whole cell lysate (ab3952) with Human FOXO3A peptide (ab27878) at 1 mg/ml
Lane 8 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate with Human FOXO3A peptide (ab27878) at 1 mg/ml
Lane 9 : 293T Cell extract - treated with Calyculin A with Human FOXO3A peptide (ab27878) at 1 mg/ml
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 89,90 kDa
Observed band size: 80 kDa (why is the actual band size different from the predicted?)
Additional bands at: 35 kDa. We are unsure as to the identity of these extra bands. -
ELISA - Anti-FOXO3A (phospho T32) antibody (ab26649)ab26649 specifically recognises the immunizing FOXO3A (phospho T32) peptide in ELISA analysis (orange dotted line), but not the corresponding unmodified peptide (yellow dotted line).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXO3A (phospho T32) antibody (ab26649)IHC image of FOXO3A staining in human Hodgkin's lymphoma FFPE section, performed on a BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab26649, 5µg/ml, for 8 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
References
ab26649 has not yet been referenced specifically in any publications.
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