製品の概要

  • 製品名
  • 製品の詳細
    Rabbit polyclonal to FOXP3
  • 特異性
    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.
  • アプリケーション
    適用あり: Flow Cyt, WB, IHC-Fr, ICC, ICC/IFmore details
    適用なし: IHC-P
  • 種交差性
    交差種: Human
    交差が予測される動物種: Macaque monkey
  • 免疫原

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

  • ポジティブ・コントロール
    • 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)
  • 特記事項
    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.

製品の特性

アプリケーション

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.

アプリケーション Abreviews 特記事項
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.
  • 追加情報
    Is unsuitable for IHC-P.
  • ターゲット情報

    • 機能
      Probable transcription factor. Plays a critical role in the control of immune response.
    • 関連疾患
      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.
    • 配列類似性
      Contains 1 C2H2-type zinc finger.
      Contains 1 fork-head DNA-binding domain.
    • 細胞内局在
      Nucleus.
    • Information by UniProt
    • 参照データベース
    • 別名
      • 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

    画像

    • 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.

    参考文献

    This product has been referenced in:
    • Tyagi RK  et al. Human IDO-competent, long-lived immunoregulatory dendritic cells induced by intracellular pathogen, and their fate in humanized mice. Sci Rep 7:41083 (2017). Read more (PubMed: 28198424) »
    • Zhu H  et al. Human Embryonic Stem Cell Lines with Lesions in FOXP3 and NF1. PLoS One 11:e0151836 (2016). IF . Read more (PubMed: 26990425) »

    See all 33 Publications for this product

    レビューと Q&A

    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

    投稿 Sep 07 2012

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (HCT116)
    Specification
    HCT116
    Fixative
    Acetone
    Permeabilization
    Yes - PBST
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C
    Username

    Abcam user community

    Verified customer

    投稿 Oct 04 2010

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Human Tissue sections (gut)
    Specification
    gut
    Fixative
    Acetone
    Permeabilization
    No
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 22°C
    Username

    Dr. alexandre garin

    Verified customer

    投稿 Nov 08 2007

    For use of product Anti-FOXP3 antibody (ab10563) in Flow Cytometry applications, we can recommend the following FITC-conjugated secondary Ab:

    ...

    Read More

    For use of product Anti-FOXP3 antibody (ab10563), we can recommend the following isotype control:

    Rabbit IgG, polyclonal - Isotype Control (ab37415).

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

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