Recombinant
RabMAb

Recombinant Anti-FOXO3A (phospho S253) antibody [EPR1951(2)] (ab154786)

Overview

  • Product name

    Anti-FOXO3A (phospho S253) antibody [EPR1951(2)]
    See all FOXO3A primary antibodies
  • Description

    Rabbit monoclonal [EPR1951(2)] to FOXO3A (phospho S253)
  • Host species

    Rabbit
  • Specificity

    ab154786 only detects FOXO3A phosphorylated at Serine 253.
  • Tested applications

    Suitable for: WB, IHC-P, Dot blotmore details
    Unsuitable for: Flow Cyt,ICC or IP
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Phospho specific peptide corresponding to residues surrounding Serine 253 of Human FOXO3A (UniProt O43524).

  • Positive control

    • MCF-7 cell lysate treated with IGF; Human endometrial carcinoma tissue; Human pancreas tissue.
  • General notes

     

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab154786 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 1/1000 - 1/10000. Predicted molecular weight: 71 kDa.
IHC-P 1/50 - 1/100.
Dot blot Use at an assay dependent concentration.
  • Application notes
    Is unsuitable for Flow Cyt,ICC or IP.
  • 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 [EPR1951(2)] (ab154786) at 1/5000 dilution

      Lane 1 : MCF-7 (Human breast adenocarcinoma epithelial cell) whole cell lysates
      Lane 2 : MCF-7 (Human breast adenocarcinoma epithelial cell) treated with IGF-1. Whole cell lysates

      Lysates/proteins at 15 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size: 71 kDa
      Observed band size: 90 kDa
      why is the actual band size different from the predicted?


      Exposure time: 1 minute


      Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    • Immunohistochemical analysis of paraffin-embedded Human endometrial carcinoma tissue labeling FOXO3A (phospho S253) with ab154786 at 1/50 dilution.
    • Immunohistochemical analysis of paraffin-embedded Human normal pancreas tissue labeling FOXO3A (phospho S253) with ab154786 at 1/50 dilution.
    • Dot Blot analysis of Lane 1: FOXO3A (pS253) phospho peptide and Lane 2: FOXO3A non-phospho peptide with ab154786 at 1/1000 dilution. 5% NFDM/TBST was used as the blocking and diluting buffer and concentration. ab97051, Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated was used as the secondary antibody at 1/100000 dilution. Exposure time: 3 minutes.

    References

    This product has been referenced in:

    • Subbannayya T  et al. PIM1 kinase promotes gallbladder cancer cell proliferation via inhibition of proline-rich Akt substrate of 40 kDa (PRAS40). J Cell Commun Signal N/A:N/A (2019). Read more (PubMed: 30666556) »
    • Kim J  et al. Syringaresinol Reverses Age-Related Skin Atrophy by Suppressing FoxO3a-Mediated Matrix Metalloproteinase-2 Activation in Copper/Zinc Superoxide Dismutase-Deficient Mice. J Invest Dermatol 139:648-655 (2019). Read more (PubMed: 30798853) »
    See all 12 Publications for this product

    Customer reviews and Q&As

    1-3 of 3 Abreviews or Q&A

    Application
    Western blot
    Sample
    Cow Cell lysate - whole cell (VVEC (vasorum endothelial cells))
    Gel Running Conditions
    Reduced Denaturing (10)
    Loading amount
    40 µg
    Treatment
    100 uM ATP 0 5 30 min
    Specification
    VVEC (vasorum endothelial cells)
    Blocking step
    BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: RT°C

    Abcam user community

    Verified customer

    Submitted Jun 03 2016

    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
    Application
    Western blot
    Loading amount
    10 µg
    Gel Running Conditions
    Reduced Denaturing
    Sample
    Human Cell lysate - whole cell (MCF7)
    Specification
    MCF7
    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 Feb 13 2015

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com

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