製品の概要

  • 製品名
    Anti-FOXO3A antibody - ChIP Grade
    FOXO3A 一次抗体 製品一覧
  • 製品の詳細
    Rabbit polyclonal to FOXO3A - ChIP Grade
  • 特異性
    This antibody was tested in FOXO3A transfected 293T cells and has not been tested against endogenous FOX03A.
  • アプリケーション
    適用あり: IHC-P, IP, IHC-FoFr, WB, ICC/IF, ChIPmore details
  • 種交差性
    交差種: Mouse, Human, Pig
    交差が予測される動物種: Rat
  • 免疫原

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

  • ポジティブ・コントロール
    • FOXO3A transfected 293T cells
  • 特記事項
    Storage 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.

製品の特性

アプリケーション

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.

アプリケーション Abreviews 特記事項
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.

ターゲット情報

  • 機能
    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'.
  • 組織特異性
    Ubiquitous.
  • 関連疾患
    Note=A chromosomal aberration involving FOXO3 is found in secondary acute leukemias. Translocation t(6;11)(q21;q23) with MLL/HRX.
  • 配列類似性
    Contains 1 fork-head DNA-binding domain.
  • 翻訳後修飾
    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.
  • 細胞内局在
    Cytoplasm > cytosol. Nucleus. Translocates to the nucleus upon oxidative stress and in the absence of survival factors.
  • Information by UniProt
  • 参照データベース
  • 別名
    • 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 画像

  • Pig coronary artery endothelial cells were cross-linked for 10 minutes at room temperature by adding 37% formaldehyde to the culture medium. The fixed cells were lysed and the chromatin was isolated. For ChIP, sheared chromatin was incubated with Anti-FOXO3A antibody - ChIP Grade (ab12162). The cells were incubated with wortmannin (30 nM), SB 203580 (10 µM) and SP 600125 (30 µM) for 30 minutes before the addition of CGJ for 30 minutes. n = 4 different experiments. *P<0.05 versus control, #P<0.05 versus CGJ treatment. Endothelial cells were exposed to CGJ for 30 minutes and then chromatin-bound DNA was immunoprecipitated with an antibody against FoxO3a.

  • 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

  • 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

  • Anti-FOXO3A antibody - ChIP Grade (ab12162) at 1/1500 dilution + Mouse muscle nuclear cell lysate at 40 µg

    Secondary
    Donkey anti-rabbit HRP conjugate (undiluted)
    Developed using the ECL technique

    Predicted band size : 71 kDa
    Observed band size : 75 kDa (why is the actual band size different from the predicted?)


    Exposure time : 1 minute

    This image is courtesy of an anonymous abreview.

    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.
  • 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

Anti-FOXO3A antibody - ChIP Grade (ab12162) 使用論文

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) »
  • Wang XX  et al. SIRT6 protects cardiomyocytes against ischemia/reperfusion injury by augmenting FoxO3a-dependent antioxidant defense mechanisms. Basic Res Cardiol 111:13 (2016). Read more (PubMed: 26786260) »

See all 10 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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投稿 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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投稿 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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投稿 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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投稿 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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投稿 Aug 09 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
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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投稿 Jul 05 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
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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投稿 Jul 02 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
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投稿 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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投稿 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"

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