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  1. Link

    foxo3a-phospho-s253-antibody-ab47285.pdf

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Epigenetics and Nuclear Signaling Transcription Domain Families Forkhead Box
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Anti-FOXO3A (phospho S253) antibody (ab47285)

  • Datasheet
  • SDS
Reviews (8)Q&A (4)References (32)

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Western blot - Anti-FOXO3A (phospho S253) antibody (ab47285)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXO3A (phospho S253) antibody (ab47285)
  • Immunocytochemistry/ Immunofluorescence - Anti-FOXO3A (phospho S253) antibody (ab47285)
  • Immunocytochemistry/ Immunofluorescence - Anti-FOXO3A (phospho S253) antibody (ab47285)

Key features and details

  • Rabbit polyclonal to FOXO3A (phospho S253)
  • Suitable for: WB, IHC-P, ICC/IF
  • Reacts with: Mouse, Human
  • Isotype: IgG

You may also be interested in

Protein
Product image
Recombinant Human FOXO3A protein (ab114191)
Secondary
Product image
Goat Anti-Rabbit IgG H&L (HRP) (ab205718)

View more associated products

Overview

  • Product name

    Anti-FOXO3A (phospho S253) antibody
    See all FOXO3A primary antibodies
  • Description

    Rabbit polyclonal to FOXO3A (phospho S253)
  • Host species

    Rabbit
  • Specificity

    This antibody is specific for FOXO3A, only when phosphorylated at serine 253.
  • Tested applications

    Suitable for: WB, IHC-P, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Human
  • Immunogen

    Synthetic peptide corresponding to Human FOXO3A.

  • 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

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • Storage buffer

    pH: 7
    Preservative: 0.02% Sodium azide
    Constituents: PBS, 50% Glycerol, 0.87% Sodium chloride
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Purification notes

    The antibody was affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific phosphopeptide. The antibody against non-phosphopeptide was removed by chromatography using non-phosphopeptide corresponding to the phosphorylation site.
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

    • Epigenetics and Nuclear Signaling
    • Transcription
    • Domain Families
    • Forkhead Box
    • Epigenetics and Nuclear Signaling
    • Transcription
    • Domain Families
    • Forkhead Box
    • FOXO

Associated products

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Positive Controls

    • NIH 3T3 nuclear extract lysate (ab14874)
  • Recombinant Protein

    • Recombinant Human FOXO3A protein (ab114191)

Applications

Our Abpromise guarantee covers the use of ab47285 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 1/500 - 1/1000. Detects a band of approximately 67, 105 kDa (predicted molecular weight: 71 kDa).
IHC-P Use at an assay dependent concentration.
ICC/IF Use a concentration of 1 µg/ml.

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.
  • Target information above from: UniProt accession O43524 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 2309 Human
    • Entrez Gene: 56484 Mouse
    • Omim: 602681 Human
    • SwissProt: O43524 Human
    • SwissProt: Q9WVH4 Mouse
    • Unigene: 220950 Human
    • Unigene: 338613 Mouse
    • 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

    • Western blot - Anti-FOXO3A (phospho S253) antibody (ab47285)
      Western blot - Anti-FOXO3A (phospho S253) antibody (ab47285)
      All lanes : Anti-FOXO3A (phospho S253) antibody (ab47285) at 1/500 dilution

      Lane 1 : extracts from NIH/3T3 cells untreated
      Lane 2 : extracts from NIH/3T3 cells treated with serum

      Lysates/proteins at 30 µg per lane.

      Predicted band size: 71 kDa
      Observed band size: 105,67 kDa
      why is the actual band size different from the predicted?



      Serum treatment:
        1. Add fresh basic medium to the vessels and starve the cells for 18-24 hours.
        2. Drain the cultured medium, add growth medium and basic medium containing 10% serum to the vessels.
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXO3A (phospho S253) antibody (ab47285)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXO3A (phospho S253) antibody (ab47285)
      Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using ab47285 at a 1/50 dilution.

      Left image : Untreated

      Right image : Treated with P-Peptide

    • Immunocytochemistry/ Immunofluorescence - Anti-FOXO3A (phospho S253) antibody (ab47285)
      Immunocytochemistry/ Immunofluorescence - Anti-FOXO3A (phospho S253) antibody (ab47285)
      ICC/IF image of ab47285 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab47285, 1µg/ml) overnight at +4°C. 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) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

    • Immunocytochemistry/ Immunofluorescence - Anti-FOXO3A (phospho S253) antibody (ab47285)
      Immunocytochemistry/ Immunofluorescence - Anti-FOXO3A (phospho S253) antibody (ab47285)This image is courtesy of an anonymous Abreview.

      ab47285 at 1/200 dilution staining FOXO3A in human 293FT cells by Immunocytochemistry/ Immunofluorescence. Cells were fixed in formaldehyde, permebilized in 0.5% Triton X-100 and then blocked in 5% BSA for 1 hour at 25°C. The primary antibody was used at 1/200 dilution in PBS and incubated with sample at 4°C for 12 hours. An Alexa Fluor® 488 conjugated Goat polyclonal to rabbit IgG was used as secondary at 1/500 dilution. Image show green staining with ab47285 and blue with DAPI.

      See Abreview

    Protocols

    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols
    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet
    • SDS
  • References (32)

    Publishing research using ab47285? Please let us know so that we can cite the reference in this datasheet.

    ab47285 has been referenced in 32 publications.

    • Lallemand F  et al. The high protein expression of FOXO3, but not that of FOXO1, is associated with markers of good prognosis. Sci Rep 10:6920 (2020). PubMed: 32332845
    • Lee E  et al. Activated pathogenic Th17 lymphocytes induce hypertension following high-fructose intake in Dahl salt-sensitive but not Dahl salt-resistant rats. Dis Model Mech 13:N/A (2020). PubMed: 32179549
    • Li J  et al. Circular RNA circFBXO11 modulates hepatocellular carcinoma progress and oxaliplatin resistance through miR-605/FOXO3/ABCB1 axis. J Cell Mol Med 24:5152-5161 (2020). PubMed: 32222024
    • Xie T  et al. microRNA-582 Potentiates Liver and Lung Metastasis of Gastric Carcinoma Cells Through the FOXO3-Mediated PI3K/Akt/Snail Pathway. Cancer Manag Res 12:5201-5212 (2020). PubMed: 32636681
    • Lee JW  et al. TLR4 (toll-like receptor 4) activation suppresses autophagy through inhibition of FOXO3 and impairs phagocytic capacity of microglia. Autophagy 15:753-770 (2019). PubMed: 30523761
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-10 of 12 Abreviews or Q&A

    Question


    Inquiry: trying to purchase p-FOXO3a (Ser253)antibody. There are several antibodies for FOXO3a (ab-47285, ab31109, ab154786). They detected different different size of FOXO3a-ser253 protein. Why?

    Read More

    Abcam community

    Verified customer

    Asked on Jun 09 2015

    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.


    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Read More

    Heather Allen

    Abcam Scientific Support

    Answered on Jun 09 2015

    Western blot abreview for Anti-FOXO3A (phospho S253) antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Loading amount
    30 µg
    Gel Running Conditions
    Reduced Denaturing (10)
    Sample
    African Green Monkey Cell lysate - whole cell (CV-1 cell kidney epithel)
    Specification
    CV-1 cell kidney epithel
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
    Read More

    Abcam user community

    Verified customer

    Submitted Nov 26 2013

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-FOXO3A (phospho S253) antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (colon carcinoma)
    Specification
    colon carcinoma
    Fixative
    Formaldehyde
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: citrate buffer (pH 6.0)
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 24°C
    Read More

    Abcam user community

    Verified customer

    Submitted Feb 05 2013

    Question

    Alternativ forab47285 Anti-FOXO3A (phospho S253) antibody, as lead time is very long. Must be tested formouse, human; IHC-P,-Fr and WB.

    Read More

    Abcam community

    Verified customer

    Asked on Mar 22 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

    Abcam Scientific Support

    Answered on Mar 22 2012

    Question

      Thanks for Support. Please find below replies to the questions you have. Hope they may assist. ab47409 - Anti-FOXO3A antibody- Order No: 946985 (1023048) - for Steve 3.1 Concentration: 1:500 Question 3.1 Have you tried different dilations such as 1/000?   yes, but 1/1000 was not as good as 1/500. 3.2 Secondary antibody: rabbit Question 3.2 Could you please specify the host species of the secondary antibody used and what type of immunoglobulin it is specific to? rabbit   IgG  3.3 Signal enhancing detection: Question 3.3 This antibody has been used for ICC/IF at 1 or 5 ug/ml using signal enhancing detection system (polymer-base   The point is the antibody does not label the phoph-foxo3 that we have confirm exist in the cytoplasm with the antibody of p253-foxo3.      

    Read More

    Abcam community

    Verified customer

    Asked on Dec 01 2011

    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

    Abcam Scientific Support

    Answered on Dec 01 2011

    Question

    Dear technical support: This customer has highly interested in ab11981 (anti-JunB) antibody), ab30628(anti-JunB (phospho S259) antibody), ab7963 (anti-c-Fos antibody), ab47285 (anti-FOXO3A (phospho S253) antibody), ab47409(anti-FOXO3A antibody), ab2732(anti-mTOR antibody), ab51044 (anti-mTOR (phospho S2448) antibody) and ab36532 (anti-IGF1 antibody), he would likes to order these antibodies and test in WB with Zebrafish sample, would you please help this customer to create a discount code, the information of this customer as follow:    Could you please help this customer to solve the problem?   Thanks for your kindly help.   Best regards  

    Read More

    Abcam community

    Verified customer

    Asked on Nov 18 2011

    Answer

    I am sending you 4 discount codes at the moment. Please try the following antibodies and let us know the results. I am happy in creating discount codes for rest of the antibodies next month. DISCOUNT CODE: XXXXXXX Expiration date: 17 March 2012 I am very pleased to hear you would like to accept our offer and test ab11981, ab30628, ab41136 ans ab42351 in Zebrafish. This code will give you 1 free [PRIMARY ANTIBODY before the expiration date. To redeem this offer, please submit an Abreview for Zebrafish and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews. Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code. Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research. The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.  

    Read More

    Abcam Scientific Support

    Answered on Nov 18 2011

    Western blot abreview for Anti-FOXO3A (phospho S253) antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Rat Cell lysate - whole cell (rat vscular smooth muscle cells)
    Loading amount
    10 µg
    Specification
    rat vscular smooth muscle cells
    Gel Running Conditions
    Reduced Denaturing
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
    Read More

    Abcam user community

    Verified customer

    Submitted Oct 04 2011

    Immunocytochemistry/ Immunofluorescence abreview for Anti-FOXO3A (phospho S253) antibody

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Rat Cell (Bone marrow cells)
    Specification
    Bone marrow cells
    Fixative
    Methanol
    Permeabilization
    No
    Blocking step
    BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    Submitted Dec 09 2010

    Western blot abreview for Anti-FOXO3A (phospho S253) antibody

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (293 FT cell line)
    Loading amount
    50 µg
    Specification
    293 FT cell line
    Gel Running Conditions
    Reduced Denaturing
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    Submitted Aug 25 2010

    Immunocytochemistry/ Immunofluorescence abreview for Anti-FOXO3A (phospho S253) antibody

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (293FT cell line)
    Specification
    293FT cell line
    Fixative
    Formaldehyde
    Permeabilization
    Yes - 0.5% Triton X-100
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    Submitted Jan 22 2010

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