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-Fr, IHC-P, ELISA, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide corresponding to Human FOXO3A.

  • Positive control
    • Human breast carcinoma tissue; NIH/3T3 cell extracts

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • Storage buffer
    pH: 7.40
    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

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-Fr Use at an assay dependent concentration. See Abreview.
IHC-P Use at an assay dependent concentration.
ELISA 1/20000.
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.
  • 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

  • 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.
  • 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

  • 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.

  • 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

References

This product has been referenced in:
  • Zhang H  et al. The mechanisms involved in miR-9 regulated apoptosis in cervical cancer by targeting FOXO3. Biomed Pharmacother 102:626-632 (2018). Read more (PubMed: 29602130) »
  • Liu S  et al. FoxO3a plays a key role in the protective effects of pomegranate peel extract against amikacin-induced ototoxicity. Int J Mol Med 40:175-181 (2017). WB, IHC . Read more (PubMed: 28560451) »
See all 16 Publications for this product

Customer reviews and Q&As

1-10 of 12 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.


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

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

Abcam user community

Verified customer

Submitted Nov 26 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
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

Abcam user community

Verified customer

Submitted Feb 05 2013

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

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 guarantees this product to work in the species/application used in this Abreview.
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

Abcam user community

Verified customer

Submitted Oct 04 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
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

Abcam user community

Verified customer

Submitted Dec 09 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
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

Abcam user community

Verified customer

Submitted Aug 25 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
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

Abcam user community

Verified customer

Submitted Jan 22 2010

1-10 of 12 Abreviews or Q&A

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