Anti-FOXO3A (phospho T32) antibody (ab26649)

Overview

  • Product nameAnti-FOXO3A (phospho T32) antibody
    See all FOXO3A primary antibodies
  • Description
    Rabbit polyclonal to FOXO3A (phospho T32)
  • Tested applicationsSuitable for: WB, IHC-Pmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Mouse, Chicken, Xenopus laevis, Zebrafish
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human FOXO3A, phosphorylated at T32.

    (Peptide available as ab27877.)

  • Positive control
    • This antibody gave a positive signal in the following whole cell lysates; HT29; Jurkat; 293T Cell extract - treated with Calyculin A,

Properties

Associated products

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

  • 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'.
  • Tissue specificityUbiquitous.
  • 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.
  • Post-translational
    modifications
    In 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
  • Database links
  • Alternative names
    • AF6q21 antibody
    • AF6q21 protein antibody
    • DKFZp781A0677 antibody
    • FKHR2 antibody
    • FKHRL 1 antibody
    • FKHRL1 antibody
    • FKHRL1P2 antibody
    • Forkhead (Drosophila) homolog (rhabdomyosarcoma) like 1 antibody
    • Forkhead box O3 antibody
    • Forkhead box O3A antibody
    • Forkhead box protein O3 antibody
    • Forkhead box protein O3A antibody
    • Forkhead Drosophila homolog of in rhabdomyosarcoma like 1 antibody
    • Forkhead homolog (rhabdomyosarcoma) like 1 antibody
    • Forkhead in rhabdomyosarcoma like 1 antibody
    • Forkhead in rhabdomyosarcoma-like 1 antibody
    • FOX O3A antibody
    • FOXO2 antibody
    • foxo3 antibody
    • FOXO3_HUMAN antibody
    • FOXO3A antibody
    • MGC12739 antibody
    • MGC31925 antibody
    see all

Anti-FOXO3A (phospho T32) antibody images

  • 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 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 : 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
    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.
  •  ab26649 specifically recognises the immunizing FOXO3A (phospho T32) peptide in ELISA analysis (orange dotted line), but not the corresponding unmodified peptide (yellow dotted line).

        
  • 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 for Anti-FOXO3A (phospho T32) antibody (ab26649)

ab26649 has not yet been referenced specifically in any publications.

Product Wall

Application Western blot
Loading amount 30 µg
Gel Running Conditions Non-reduced Denaturing (8% gel)
Sample Pig Cell lysate - whole cell (Endothelial Cell)
Specification Endothelial Cell
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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Verified customer

Submitted Sep 02 2014

Thank you for getting back to me and for providing some further information. It is much appreciated. This antibody is rabbit polyclonal so the host species of the compatible secondary antibody should be different from rabbit. Reading your respo...

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Application Western blot
Sample Human Cell lysate - whole cell (Calu 6 (Lung))
Loading amount 40 µg
Specification Calu 6 (Lung)
Treatment IGF1 (10nM) Time (0, 5, 15 min)
Gel Running Conditions Reduced Denaturing
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Username

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Verified customer

Submitted Sep 30 2010

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"