Anti-FOXO3A (phospho S253) 抗体 (ab47285)

製品の概要

  • 製品名Anti-FOXO3A (phospho S253) antibody
    FOXO3A 一次抗体 製品一覧
  • 製品の詳細
    Rabbit polyclonal to FOXO3A (phospho S253)
  • 特異性This antibody is specific for FOXO3A, only when phosphorylated at serine 253.
  • アプリケーション適用あり: WB, IHC-Fr, IHC-P, ELISA, ICC/IFmore details
  • 種交差性
    交差種: Mouse, Rat, Human
  • 免疫原

    A synthesized phosphopeptide derived from human FOXO3A around the phosphorylation site of serine 253 (A-V-SP-M-D)

  • ポジティブ・コントロール
    • Human breast carcinoma tissue; NIH/3T3 cell extracts

製品の特性

  • 製品の状態Liquid
  • 保存方法Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • バッファーPreservative: 0.02% Sodium Azide
    Constituents: 50% Glycerol, PBS, 150mM Sodium chloride, pH 7.4
  • Concentration information loading...
  • 精製度Immunogen affinity purified
  • 特記事項(精製)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.
  • ポリ/モノポリクローナル
  • アイソタイプIgG
  • 研究分野

アプリケーション

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.

アプリケーション Abreviews 特記事項
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.

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 (phospho S253) antibody 画像

  • 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 : 67,105 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

Anti-FOXO3A (phospho S253) antibody (ab47285) 使用論文

This product has been referenced in:
  • Holloway TM  et al. High intensity interval and endurance training have opposing effects on markers of heart failure and cardiac remodeling in hypertensive rats. PLoS One 10:e0121138 (2015). WB ; Rat . Read more (PubMed: 25803693) »
  • Fiskus W  et al. Dual PI3K/AKT/mTOR inhibitor BEZ235 synergistically enhances the activity of JAK2 inhibitor against cultured and primary human myeloproliferative neoplasm cells. Mol Cancer Ther 12:577-88 (2013). Read more (PubMed: 23445613) »

See all 6 Publications for this product

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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 th...

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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
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投稿 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
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投稿 Feb 05 2013

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 mö...

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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 respo...

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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 ...

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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
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投稿 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
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投稿 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
Username

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Verified customer

投稿 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
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投稿 Jan 22 2010

1-10 of 12 Abreviews or Q&A

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