概述

  • 产品名称Anti-FOXP3抗体
    参阅全部 FOXP3 一抗
  • 描述
    兔多克隆抗体to FOXP3
  • 特异性This antibody recognises a single clean band in a HEK293 cell lysate that overexpresses human FOXP3. The band is not seen in untransfected cells and is blocked by the immunising peptide. This antibody also recognises a band at 50 kDa in Human thymus, spleen and tonsil tissue extracts.
  • 经测试应用适用于: Flow Cyt, WB, IHC-Fr, ICC, ICC/IFmore details
    不适用于: IHC-P
  • 种属反应性
    与反应: Human
    预测可用于: Macaque Monkey
  • 免疫原

    Synthetic peptide corresponding to Human FOXP3 aa 400-500 conjugated to Keyhole Limpet Haemocyanin (KLH).
    (Peptide available as ab14151)

  • 阳性对照
    • ICC: U2OS cells WB: HEK 293 lysate over expressing human FOXP3, Human thymus, tonsil, spleen tissue IHC-Fr: human tonsil FACS: purified human CD4+, CD4+CD25- and CD4+CD25+ cells (see review)
  • 常规说明There is a reported difference in the human and mouse immune systems regarding FOXP3 expression being induced by T cell activation. In contrast to the murine system, the activation of human CD4+ CD25- T cells through TCR leads to the increased expression of FOXP3 mRNA, the surface expression of CD25 and gain of regulatory function (see references especially Walker et al, 2003). This suggests that FOXP3 in human is not a unique marker for regulatory T cells. The pattern of staining reported with ab10563 is consistent with the pattern of other anti-FOXP3 antibodies commercially available and we would welcome your opinion regarding the difference in population stained with FOXP3 antibodies; certain users have indeed observed a staining in human CD25- cells as reported in several papers.

性能

应用

Our Abpromise guarantee covers the use of ab10563 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
Flow Cyt Use a concentration of 4 µg/ml. ab171870-Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.
WB Use a concentration of 1 - 5 µg/ml. Detects a band of approximately 50 kDa (predicted molecular weight: 47.2 kDa).
IHC-Fr Use a concentration of 0.625 µg/ml.
ICC Use a concentration of 2.5 - 5 µg/ml.
ICC/IF Use at an assay dependent concentration.
  • 应用说明Is unsuitable for IHC-P.
  • 靶标

    • 功能Probable transcription factor. Plays a critical role in the control of immune response.
    • 疾病相关Defects in FOXP3 are the cause of immunodeficiency polyendocrinopathy, enteropathy, X-linked syndrome (IPEX) [MIM:304790]; also known as X-linked autoimmunity-immunodeficiency syndrome. IPEX is characterized by neonatal onset insulin-dependent diabetes mellitus, infections, secretory diarrhea, trombocytopenia, anemia and eczema. It is usually lethal in infancy.
    • 序列相似性Contains 1 C2H2-type zinc finger.
      Contains 1 fork-head DNA-binding domain.
    • 细胞定位Nucleus.
    • Information by UniProt
    • 数据库链接
    • 别名
      • AIID antibody
      • DIETER antibody
      • Forkhead box P3 antibody
      • Forkhead box protein P3 antibody
      • FOXP3 antibody
      • FOXP3_HUMAN antibody
      • FOXP3delta7 antibody
      • Immune dysregulation polyendocrinopathy enteropathy X linked antibody
      • Immunodeficiency polyendocrinopathy enteropathy X linked antibody
      • IPEX antibody
      • JM2 antibody
      • MGC141961 antibody
      • MGC141963 antibody
      • OTTHUMP00000025832 antibody
      • OTTHUMP00000025833 antibody
      • OTTHUMP00000226737 antibody
      • PIDX antibody
      • Scurfin antibody
      • XPID antibody
      see all

    Anti-FOXP3 antibody 图像

    • FoxP3 immunostaining on a frozen section from human tonsil. Original magnification x100. Nuclear staining was observed.
    • Lanes 1 - 6 : Anti-FOXP3 antibody (ab10563) at 1/500 dilution
      Lane 7 : No-primary control

      Lane 1 : 20µg HEK 293 lysate
      Lane 2 : 20µg HEK 293 lysate over expressing human FOXP3
      Lane 3 : 20µg HEK 293 lysate over expressing mouse FOXP3
      Lane 4 : 20µg HEK 293 lysate with Human FOXP3 peptide (ab14151) at 1 µg/ml
      Lane 5 : 20µg HEK 293 lysate over expressing human FOXP3 with Human FOXP3 peptide (ab14151) at 1 µg/ml
      Lane 6 : 20µg HEK 293 lysate over expressing mouse FOXP3 with Human FOXP3 peptide (ab14151) at 1 µg/ml
      Lane 7 : 20µg HEK 293 lysate

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab6721) at 1/5000 dilution

      Performed under reducing conditions.

      Predicted band size : 47.2 kDa
      Observed band size : 47 kDa (why is the actual band size different from the predicted?)


      Exposure time : 30 seconds

      This antibody recognises a single clean band in a HEK293 cell lysate that overexpresses human FOXP3. The band is not seen in untransfected cells and is blocked by the immunising peptide.

    • Immunofluorescent imaging of human cells (U2OS) with ab10563 (5 ug/ml) confirms the specificity of this anti-FOXP3 antibody, which recognises nuclear signal only; with no background cytoplasmic or nucleolar staining.  

      IF was performed with a standard paraformaldehyde technique (fixed in PBS buffered PFH 4% for 5 minutes, permeabilised with 0.5% triton-PBS for 5 minutes, blocked with 5% milk / 0.2% tween for one hour).  Primary antibody used at 5ug/ml in 5% milk / 0.2% TWEEN for 1 hour, secondary antibody for 30 minutes.  All blocking and incubation steps carried out at 37 degrees C. Nuclei were stained with Hoechst stain.

    • All lanes : Anti-FOXP3 antibody (ab10563) at 5 µg/ml

      Lane 1 : Human thymus tissue lysate
      Lane 2 : Human spleen tissue lysate
      Lane 3 : Human tonsil tissue lysate

      Lysates/proteins at 20 µg per lane.


      Performed under reducing conditions.

      Predicted band size : 47.2 kDa

      This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using Licor blocking buffer before being incubated with ab10563. Antibody binding was detected using Goat anti Rabbit IR680 at a 1:10,000 dilution for 1hr at room temperature and then imaged using the Licor Odyssey CLx.

    Anti-FOXP3 antibody (ab10563)参考文献

    This product has been referenced in:
    • Tan B  et al. FOXP3 over-expression inhibits melanoma tumorigenesis via effects on proliferation and apoptosis. Oncotarget 5:264-76 (2014). IF . Read more (PubMed: 24406338) »
    • Xiao F  et al. Ex vivo expanded human regulatory T cells delay islet allograft rejection via inhibiting islet-derived monocyte chemoattractant protein-1 production in CD34+ stem cells-reconstituted NOD-scid IL2r?null mice. PLoS One 9:e90387 (2014). IHC-P . Read more (PubMed: 24594640) »

    See all 22 Publications for this product

    Product Wall

    Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.
    I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has c...

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    Because we carry over 90,000 products, it isn't feasible for us to keep small sample sizes of our products, and therefore I am not able to offer a test sample of ab20034 or ab10563.

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    Herr K. hat mich darüber informiert, dass Sie unsere ab10563und ab78007 (FOXP3 und Anti-ROR gamma Antikörper) gerne in der IHC am Katzengewebetesten möchten. Leider ist die Homologie zwischen dem humanen und felinem ROR ga...

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    Thank you for contacting us.

    There isn’t a specific protocol for performing IHC-P using this antibody. According to the datasheet the antibody concentration is assay dependant, and optimisation is recommended by using different antibody...

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    Abcam has not validated the combination of species/application used in this Abreview.
    Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample Human Tissue sections (Tonsil)
    Specification Tonsil
    Fixative Formaldehyde
    Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citric acid
    Permeabilization No
    Blocking step BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C
    Username

    Mr. Carl Hobbs

    Verified customer

    提交于 Sep 07 2012

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    Your credit note ID is 22440.

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    I have attached the filled out Exel spreadsheet with all the requested information. To be fair, all the information asked could be answered with a look on the datasheet...

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    I am sorry that this antibody did not perform as stated on the datasheet, I have asked our Finance department to issue a credit note for you. The credit note may be used i...

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    Vielen Dank für Ihre Anfrage.

    Ich habe Ihnen hier vier Antikörper herausgesucht, die wenig bzw. gar keine Kreuzreaktion mit dem Maus-Protein aufweisen, so dass Sie nur das humane Protein im Mausgewebe detektieren sollten.

    Drei der...

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    Thank you for contacting us.

    In order to understand the problem I would like toask following queations;

    - In the attached details it is mentioned that Rabbit secondary was used. Could you please double check because rabbit secondar...

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