Overview

  • Product nameAnti-FOXO3A antibody - ChIP Grade
    See all FOXO3A primary antibodies
  • Description
    Rabbit polyclonal to FOXO3A - ChIP Grade
  • SpecificityThis antibody was tested in FOXO3A transfected 293T cells and has not been tested against endogenous FOX03A.
  • Tested applicationsSuitable for: IHC-P, IP, IHC-FoFr, WB, ICC/IF, ChIPmore details
  • Species reactivity
    Reacts with: Mouse, Human, Pig
    Predicted to work with: Rat
  • Immunogen

    Synthetic peptide: C-GLNVGNFTGAKQASSQ, with N-terminally added cysteine, conjugated to maleimide activated KLH, corresponding to amino acids 653-668 of Human FOXO3A.

  • Positive control
    • FOXO3A transfected 293T cells
  • General notesStorage in frost-free freezers is not recommended. If slight turbidity occurs upon prolonged storage, clarify the solution by centrifugation before use. Working dilutions should be discarded if not used within 12 hours.

Properties

Applications

Our Abpromise guarantee covers the use of ab12162 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
IHC-P Use at an assay dependent concentration.
IP 1/600.
IHC-FoFr 1/300.
WB 1/2500. Detects a band of approximately 70-75 kDa (predicted molecular weight: 71 kDa). In some preparations there is an additional band at 35 kDa.
A customer reported the antibody did not detect endogenous FOX03A in human cancer cells (see enquiries).
ICC/IF Use at an assay dependent concentration.
ChIP Use at an assay dependent concentration.

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 antibody - ChIP Grade images

  • ab12162 staining FOXO3A in Rat fibroblast cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.03% Triton X-100 and blocked with 5% serum for 10 minutes at 25°C. Samples were incubated with primary antibody (1/300) for 1 hour at 25°C.

    See Abreview



  • Predicted band size : 71 kDa

    Image from Åhsberg J et al, J Biol Chem. 2010 Nov 19;285(47):36275-84. Epub 2010 Sep 9, Fig 7. DOI 10.1074/jbc.M110.155531 November 19, 2010 Journal of Biological Chemistry, 285, 36275-36284.

    For protein extraction, murine BaF3Flt-3,Il-7 cells were washed in ice-cold PBS and immediately resuspended in lysis buffer containing 25 mm Tris (pH 7.5), 1% Triton X-100, 150 mm NaCl, 1 mm DTT, 1 mm EDTA (pH 8.0), 1× protease inhibitor mixture and 1 mm Na3VO4. Samples were centrifuged 1000 × g for 2 minutes and supernatant was mixed with 1× LDS sample buffer and 0.01 m DTT. Samples were heated at 70 °C for 10 minutes before separated by SDS-PAGE and blotted onto a PVDF transfer membrane. Membranes were blocked in 5% milk PBS/Tween or 5% BSA TBS/Tween for 1 hour at room temperature and next incubated with primary antibodies for 1 hour at room temperature or at 8 °C overnight. Following block, membranes were incubated with HRP-conjugated secondary antibody for 1 hour at room temperature. Blots were developed by enhanced chemiluminescence.
  • Immunohistochemical analysis of formalin-fixed, paraffin-embedded Human colon carcinoma tissue, staining FOXO3A with ab12162.

    Tissue was fixed with formaldehyde and blocked with 1% serum for 1 hour at 24°C; antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/100 in diluent) for 12 hours at 4°C. A biotinylated goat anti-rabbit polyclonal IgG (1/200) was used as the secondary antibody.

    See Abreview

  • ab12162 staining FOXO3A in human brain tissue sections by Immunohistochemistry (PFA perfusion fixed frozen sections). Tissue samples were fixed by perfusion with formaldehyde, permeablized with 0.01% Triton-X 100 and blocked with 5% serum for 4 hours at 25°C. The sample was incubated with primary antibody (1/300) at 4°C for 10 hours. An Alexa Fluor® 594-conjugated donkey anti-rabbit IgG polyclonal (1/300) was used as the secondary antibody.

    See Abreview

References for Anti-FOXO3A antibody - ChIP Grade (ab12162)

This product has been referenced in:
  • Liu JY  et al. MicroRNA-153 inhibits the proliferation and invasion of human laryngeal squamous cell carcinoma by targeting KLF5. Exp Ther Med 11:2503-2508 (2016). WB ; Human . Read more (PubMed: 27284339) »
  • Alhosin M  et al. Redox-sensitive up-regulation of eNOS by purple grape juice in endothelial cells: role of PI3-kinase/Akt, p38 MAPK, JNK, FoxO1 and FoxO3a. PLoS One 8:e57883 (2013). ChIP ; Pig . Read more (PubMed: 23533577) »

See all 7 Publications for this product

Product Wall

Application Immunohistochemistry (PFA perfusion fixed frozen sections)
Blocking step Serum as blocking agent for 4 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Sample Human Tissue sections (Brain)
Specification Brain
Permeabilization Yes - 0.01% Triton X100
Fixative Formaldehyde
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Submitted Jun 23 2014

Application ChIP
Detection step Semiquantitative PCR
Sample Rat Cell lysate - whole cell (Rat primary cardiomyocytes)
Specification Rat primary cardiomyocytes
Type Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: Formaldehyde
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Submitted Mar 31 2014

Application Immunocytochemistry/ Immunofluorescence
Blocking step BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 22°C
Sample Human Cell (HepG2)
Specification HepG2
Permeabilization No
Fixative Paraformaldehyde
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Submitted Feb 11 2014

Application Western blot
Loading amount 30 µg
Gel Running Conditions Reduced Denaturing
Sample Rat Cell lysate - whole cell (Glial cells)
Specification Glial cells
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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Submitted Aug 09 2013

Application Immunocytochemistry/ Immunofluorescence
Blocking step Serum as blocking agent for 10 minute(s) · Concentration: 5% · Temperature: 25°C
Sample Rat Cell (fibroblast)
Specification fibroblast
Permeabilization Yes - 0.03% TritonX-100
Fixative Paraformaldehyde
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Submitted Aug 09 2013

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Application Immunoprecipitation
Immuno-precipitation step Protein A
Sample Mouse Cell lysate - nuclear (primary hepatoctyes)
Specification primary hepatoctyes
Total protein in input 50 µg
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Submitted Jul 05 2013

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Application Western blot
Loading amount 50 µg
Gel Running Conditions Non-reduced Non-Denaturing (Native) (10%)
Sample Human Cell lysate - whole cell (HepG2)
Specification HepG2
Treatment 48 h melatonin treatment from 50 to 2000mM
Blocking step (agent) for 30 minute(s) · Concentration: 2µg/mL · Temperature: 22°C
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Submitted Jul 02 2013

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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
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Submitted Feb 05 2013

Thank you for contacting us.
I am sorry to hear you are experiencing difficulties with one of our products. We take product complaints very seriously, and investigate every product that we feel may not be performing correctly.
The ChIP applic...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (rat VSMC expression of human foxo3a conjugate prot)
Loading amount 100000 cells
Specification rat VSMC expression of human foxo3a conjugate prot
Gel Running Conditions Reduced Denaturing
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
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Submitted May 21 2012

1-10 of 16 Abreviews or Q&A

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