RabMAb

Anti-FOXO3A抗体[EP1949Y] (ab53287)

概述

  • 产品名称Anti-FOXO3A抗体[EP1949Y]
    参阅全部 FOXO3A 一抗
  • 描述
    兔单克隆抗体[EP1949Y] to FOXO3A
  • 经测试应用适用于: WB, IHC-P, ICC/IFmore details
    不适用于: Flow Cyt or IP
  • 种属反应性
    与反应: Human
  • 免疫原

    Synthetic peptide corresponding to residues near the N-terminus of human FOXO3A.

  • 阳性对照
    • Jurkat cell lysate and human breast carcinoma tissue. This antibody gave a positive result when used in the following methanol fixed cell lines: HeLa
  • 常规说明

    This product is a recombinant rabbit monoclonal antibody.

     

    Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.

     

    Alternative versions available:

    Anti-FOXO3A antibody (Alexa Fluor® 488) [EP1949Y] (ab196322)

    Anti-FOXO3A antibody (Alexa Fluor® 647) [EP1949Y] (ab196539)

    Anti-FOXO3A antibody (HRP) [EP1949Y] (ab196323)

性能

应用

Our Abpromise guarantee covers the use of ab53287 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 1/2000 - 1/5000. Detects a band of approximately 90 kDa (predicted molecular weight: 71 kDa).
IHC-P Use at an assay dependent concentration.
ICC/IF 1/50 - 1/100.
  • 应用说明Is unsuitable for Flow Cyt or IP.
  • 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 antibody [EP1949Y] 图像

    • ICC/IF image of ab53287 stained HeLa cells. The cells were 100% methanol fixed (5 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 ab53287 at 1/200 dilution overnight at +4°C. The secondary antibody (green) was goat anti-rabbit DyLight® 488 (ab96899) IgG (H+L) used at a 1/250 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.

    • Anti-FOXO3A antibody [EP1949Y] (ab53287) at 1/5000 dilution + Jurkat cell lysate at 10 µg

      Secondary
      Goat anti-Rabbit HRP labeled at 1/2000 dilution

      Predicted band size : 71 kDa
      Observed band size : 90 kDa (why is the actual band size different from the predicted?)
    • All lanes : Anti-FOXO3A antibody [EP1949Y] (ab53287) at 1/2000 dilution

      Lane 1 : HCT116 cells treated with DMSO (control) whole cell lysate
      Lane 2 : HCT116 cells treated with 2 µM SAHA treatment for 24 hours, whole cell lysate

      Lysates/proteins at 30 µg per lane.

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution
      Developed using the ECL technique

      Performed under reducing conditions.

      Predicted band size : 71 kDa
      Observed band size : 70 kDa (why is the actual band size different from the predicted?)


      Exposure time : 10 seconds

      This image is courtesy of an abreview submitted by Christian Marx

      Non-specific band : 130 kDa

      Blocking buffer: 5% milk

      Diluting buffer: 1% Milk in PBS-T buffer

      See Abreview

    • Ab53287 (1:50) staining human FOXO3A in human breast carcinoma tissue by immunohistochemistry using paraffin embedded tissue.

    Anti-FOXO3A antibody [EP1949Y] (ab53287)参考文献

    This product has been referenced in:
    • Cheung K  et al. CD31 signals confer immune privilege to the vascular endothelium. Proc Natl Acad Sci U S A 112:E5815-24 (2015). ICC/IF ; Mouse . Read more (PubMed: 26392551) »
    • Furgason JM  et al. Whole genome sequencing of glioblastoma multiforme identifies multiple structural variations involved in EGFR activation. Mutagenesis 29:341-50 (2014). Read more (PubMed: 25103728) »

    See all 6 Publications for this product

    Product Wall


    For IHC-P, the antigen retrieval solution used was citrate buffer pH6 - heat-mediated.

    Antigen Retrieval

    Immerse slides into staining dish containing Antigen Retrieval Solution.

    Place covered staining dish into the rice ...

    Read More
    Application Western blot
    Sample Mouse Cell lysate - whole cell (MEF cell line)
    Loading amount 30 µg
    Specification MEF cell line
    Treatment DMSO control and 2 µM SAHA for 24 hrs
    Gel Running Conditions Reduced Denaturing (13%)
    Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Username

    Mr. Christian Marx

    Verified customer

    提交于 Feb 22 2013

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Western blot
    Sample Human Cell lysate - whole cell (HCT116 cell line)
    Loading amount 30 µg
    Specification HCT116 cell line
    Treatment DMSO control and 2 µM SAHA treatment for 24 hrs
    Gel Running Conditions Reduced Denaturing (13%)
    Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Username

    Mr. Christian Marx

    Verified customer

    提交于 Feb 20 2013

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Western blot
    Sample Human Cell lysate - whole cell (Jurkat cells)
    Loading amount 200000 cells
    Specification Jurkat cells
    Gel Running Conditions Reduced Denaturing
    Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
    Username

    Abcam user community

    Verified customer

    提交于 Nov 24 2009

    Thank you for your enquiry. Please refer to the following ICC/IF protocols for this product. Solutions and reagents - PBST washing buffer: 1XPBS/0.1% Tween-20 (Dulbecco’s Phosphate Buffered Salts, 1X, catalog #21-031-CV from Mediatech, Inc.) ...

    Read More

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