Overview

  • Product name
    Anti-FOXO3A (phospho S253) antibody
    See all FOXO3A primary antibodies
  • Description
    Rabbit polyclonal to FOXO3A (phospho S253)
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, ICC/IF, IHC-P, IPmore details
  • Species reactivity
    Reacts with: Mouse, Rat
    Predicted to work with: Human, Pig, Zebrafish
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 200 - 300 of Human FOXO3A.

    Read Abcam's proprietary immunogen policy (Peptide available as ab27885.)

  • Positive control
    • Recombinant Human FOXO3A protein (ab114191) can be used as a positive control in WB. This antibody gave a positive signal in PC12 (Rat adrenal pheochromocytoma cell line) Cytoplasmic Lysate.

Properties

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
  • Application notes
    ICC/IF: Use at a concentration of 1 µg/ml.
    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

    • 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
      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 localization
      Cytoplasm > 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

    Images

    • 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

    • 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).
    • 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.

    References

    This product has been referenced in:
    • Zeng X  et al. Acylated and unacylated ghrelin inhibit atrophy in myotubes co-cultured with colon carcinoma cells. Oncotarget 8:72872-72885 (2017). Read more (PubMed: 29069832) »
    • Gravina GL  et al. Pharmacological treatment with inhibitors of nuclear export enhances the antitumor activity of docetaxel in human prostate cancer. Oncotarget 8:111225-111245 (2017). Read more (PubMed: 29340049) »
    See all 5 Publications for this product

    Customer reviews and Q&As

    1-4 of 4 Abreviews or Q&A

    Answer

    Thank you for contacting us.


    The predicted MW of the protein is ˜ 75 kD, based on sequence alone.


    However page 4 of paper attached showed, phosphorylation on serine 253 of FOXO3A/FKHRL1 could migrate the MW to ˜100KD for this protein.


    Read More
    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Rat Tissue sections (Tongue)
    Specification
    Tongue
    Fixative
    Bouin's solution
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Tris-EDTA pH: 9
    Permeabilization
    No
    Blocking step
    Casein in sodium carbonate buffer, pH 9,6 as blocking agent for 30 minute(s) · Concentration: 0.2% · Temperature: 22°C

    Dr. Tonny Petersen

    Verified customer

    Submitted Dec 27 2012

    Answer

    Vielen Dank für Ihren Anruf und für Ihr Interesse an unseren Produkten.

    Unseres Wissens nach wurde ab31109 Anti-FOXO3A (phospho S253) antibodybisher noch nicht inIHC-Fr (Kyro-Schnitten)getestet. Falls Sie dies selbst testen möchten, kann ich Ihnen zur Zeit ein spezielles Angebot über einen 100%igen Abreview-Rabatt anbieten. Bei diesem Angebot bekommen Sie einen Rabatt für eine zukünftige Bestellung, wenn Sie uns ein Abreview mit dem Testresultat zusenden. Der Rabatt würde den ganzen Wert von ab31109 abdecken, und gegen eine erneute Bestellung eines weiterenprimären Antikörpersvon uns verrechnet werden.

    Um von diesem Angebot profitieren zu können, folgen Sie bitte diesen Schritten:

    1.) Bestätigen Sie mir bitte, dass Sie ab31109 in IHC-Fr testen möchten.

    2.) Bitte bestellen Sie erst nach Erhalt des Rabattcodes.

    3.) Bestellen Sie den Antikörper wie üblich per Telefon, Email oder Fax

    4.) Testen Sie den Antikörper in IHC-Fr.

    5.) Teilen Sie uns das Ergebnis Ihres Tests durch ein Abreview mit und notieren Sie den Discount Code in dem Feld "additional notes"
    Unter der folgenden URL können Sie mehr über unser Abreview System erfahren:
    https://www.abcam.com/abreviews

    6.) Der Rabattcode ist nach dem Abschicken des Abreviews aktiv, und Sie können einen anderen primären Antikörpers zu dem gleichen Preis wie ab31109 bei uns bestellen (halten Sie bei der Bestellung bitte den Rabattcode und die Bestellnummer von ab31109 bereit). Bitte beachten Sie, dass der Rabattcode innerhalb von 4 Monaten nach Ausstellung eingelöst werden muss.

    Der Rabattcode wird gültig unabhängig davon, ob Ihr Ergebnis positiv oder negativ ist.

    Die Bedingungen zu unserem 100% Abreview Rabatt können Sie unter dem folgenden Link nachlesen:
    https://www.abcam.com/collaborationdiscount

    Ich hoffe, diese Informationen helfen Ihnen weiter. Falls Sie einen Rabattcode erhalten möchten oder weitere Fragen haben, zögern Sie bitte nicht, sich wieder an mich zu wenden.

    Read More

    Answer

    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 response, it seems you are expecting to detect the phospho FOXO3A (at serine 253) with ab47409 . It is important to emphasize that as the datasheet indicates the immunogen used to raise ab47409 was a synthesized non-phosphopeptide derived from human FKHRL1 around the phosphorylation site of serine 253. Therefore this antibody only detects the non-phosphopeptide FOXO3A. For your information, we have some specific antibodies against different phosphorylated FOXO3A at different positions, but these product names and their specificity state that they only recognize the modified version of FOXO3A: ab47285:Anti-FOXO3A (phospho S253) antibody:https://www.abcam.com/foxo3a-phospho-s253-antibody-ab47285.html ab31109:Anti-FOXO3A (phospho S253) antibody: https://www.abcam.com/foxo3a-phospho-s253-antibody-ab31109.html ab26649:Anti-FOXO3A (phospho T32) antibody:https://www.abcam.com/foxo3a-phospho-t32-antibody-ab26649.html I hope this helps and if I can assist further, please do not hesitate to

    Read More

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