Anti-FOXO3A (phospho S253)抗体(ab47285)

概述

  • 产品名称Anti-FOXO3A (phospho S253)抗体
    参阅全部 FOXO3A 一抗
  • 描述
    兔多克隆抗体to FOXO3A (phospho S253)
  • 特异性This antibody is specific for FOXO3A, only when phosphorylated at serine 253.
  • 经测试应用适用于: WB, IHC-Fr, IHC-P, ELISA, ICC/IFmore details
  • 种属反应性
    与反应: 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)

  • 阳性对照
    • Human breast carcinoma tissue; NIH/3T3 cell extracts

性能

  • ���式Liquid
  • 存放说明Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • 存储溶液Preservative: 0.02% Sodium Azide
    Constituents: 50% Glycerol, PBS, 150mM Sodium chloride, pH 7.4
  • Concentration information loading...
  • 纯度Immunogen affinity purified
  • 纯化说明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.
  • 克隆多克隆
  • 同种型IgG
  • 研究领域

应用

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.

应用 Ab评论 说明
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.

靶标

  • 功能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'.
  • 组织特异性Ubiquitous.
  • 疾病相关Note=A chromosomal aberration involving FOXO3 is found in secondary acute leukemias. Translocation t(6;11)(q21;q23) with MLL/HRX.
  • 序列相似性Contains 1 fork-head DNA-binding domain.
  • 翻译后修饰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.
  • 细胞定位Cytoplasm > cytosol. Nucleus. Translocates to the nucleus upon oxidative stress and in the absence of survival factors.
  • Information by UniProt
  • 数据库链接
  • 别名
    • 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 图像

  • 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)参考文献

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

Product Wall

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 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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提交于 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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提交于 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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Verified customer

提交于 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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提交于 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

Abcam user community

Verified customer

提交于 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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提交于 Jan 22 2010

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