Anti-FOXO3A (phospho T32) antibody (ab26649)
Key features and details
- Rabbit polyclonal to FOXO3A (phospho T32)
- Suitable for: ELISA, WB, IHC-P
- Reacts with: Human, Recombinant fragment
- Isotype: IgG
Overview
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Product name
Anti-FOXO3A (phospho T32) antibody
See all FOXO3A primary antibodies -
Description
Rabbit polyclonal to FOXO3A (phospho T32) -
Host species
Rabbit -
Tested applications
Suitable for: ELISA, WB, IHC-Pmore details -
Species reactivity
Reacts with: Human, Recombinant fragment
Predicted to work with: Mouse, Chicken, Xenopus laevis, Zebrafish
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Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- Recombinant Human FOXO3A protein (ab114191) can be used as a positive control in WB. This antibody gave a positive signal in the following whole cell lysates; HT29; Jurkat; 293T Cell extract - treated with Calyculin A,
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General notes
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
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.
Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.
Properties
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Form
Liquid -
Storage instructions
Shipped 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. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration
Concentration information loading...Purity
Immunogen affinity purifiedClonality
PolyclonalIsotype
IgGResearch areas
Associated products
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Compatible Secondaries
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Isotype control
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Positive Controls
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Recombinant Protein
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 ELISA Use at an assay dependent concentration. 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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Function
Transcriptional 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'. -
Tissue specificity
Ubiquitous. -
Involvement in disease
Note=A chromosomal aberration involving FOXO3 is found in secondary acute leukemias. Translocation t(6;11)(q21;q23) with MLL/HRX. -
Sequence similarities
Contains 1 fork-head DNA-binding domain. -
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 localization
Cytoplasm > 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 :HT-29 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 :HT-29 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 with Human FOXO3A (phospho T32) peptide (ab27877) at 1 mg/ml
Lane 6 : 293T Cell extract - treated with Calyculin A with Human FOXO3A (phospho T32) peptide (ab27877) at 1 mg/ml
Lane 7 :HT-29 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.
Protocols
Datasheets and documents
References (3)
ab26649 has been referenced in 3 publications.
- Xie Y et al. The proteasome activator REG? accelerates cardiac hypertrophy by declining PP2Aca-SOD2 pathway. Cell Death Differ N/A:N/A (2020). PubMed: 32424140
- Li J et al. A Discovery of a Genetic Mutation Causing Reduction of Atrogin-1 Expression in Broiler Chicken Muscle. Front Genet 10:716 (2019). PubMed: 31475031
- Blanchard Z et al. BRCA1-IRIS inactivation overcomes paclitaxel resistance in triple negative breast cancers. Breast Cancer Res 17:5 (2015). PubMed: 25583261
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 :HT-29 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 :HT-29 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 with Human FOXO3A (phospho T32) peptide (ab27877) at 1 mg/ml
Lane 6 : 293T Cell extract - treated with Calyculin A with Human FOXO3A (phospho T32) peptide (ab27877) at 1 mg/ml
Lane 7 :HT-29 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.
