Anti-FOXP3 antibody [mAbcam 450] (ab450)

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Overview

  • Product name

    Anti-FOXP3 antibody [mAbcam 450]
    See all FOXP3 primary antibodies

  • Description

    Mouse monoclonal [mAbcam 450] to FOXP3

  • Host species

    Mouse

  • Specificity

    Detects a single clean band on human FOXP3 transfected 293 cells. When used in immunocytochemistry on acetone fixed FOXP protein transfected COS cells the antibody only detects FOXP3 and not FOXP1, FOXP2 or FOXP4.

  • Tested applications

    Suitable for: IHC-Fr, IP, ICC, IHC-P, ICC/IF, WBmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Synthetic peptide corresponding to Human FOXP3 aa 400 to the C-terminus conjugated to keyhole limpet haemocyanin.
    Database link: Q9BZS1
    (Peptide available as ab16809)

  • Positive control

    • IHC-P: FFPE Human tonsil tissue sections. IHC-Fr: Human tonsil tissue sections.

  • General notes

    This antibody clone is manufactured by Abcam.

    If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

Properties

Applications

Our Abpromise guarantee covers the use of ab450 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
IHC-Fr Use a concentration of 5 µg/ml.
IP Use at an assay dependent concentration. Used at 2 µg/ml on human FOXP3 transfected COS cells (see Abreview for further information).
ICC Use at an assay dependent concentration.
IHC-P 1/50.
This clone has been tested against FOXP3 transfectants of COS-1 cells fixed with acetone and also works against formalin-fixed paraffin-embedded FOXP3 transfectants heat retrieved with pH9 Tris-EDTA buffer. It did not show positive staining with cryostat or paraffin-embedded human tonsil.
ICC/IF Use at an assay dependent concentration.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 48 kDa (predicted molecular weight: 48 kDa).Can be blocked with Human FOXP3 peptide (ab16809).

Target

  • Function

    Probable transcription factor. Plays a critical role in the control of immune response.

  • Involvement in disease

    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.

  • Sequence similarities

    Contains 1 C2H2-type zinc finger.
    Contains 1 fork-head DNA-binding domain.

  • Cellular localization

    Nucleus.
  • Information by UniProt

  • Database links

    • Alternative names

      • 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

    Images

    • Western blot - Anti-FOXP3 antibody [mAbcam 450] (ab450)
      Western blot - Anti-FOXP3 antibody [mAbcam 450] (ab450)
      All lanes : Anti-FOXP3 antibody [mAbcam 450] (ab450) at 1 µg/ml

      Lane 1 : HEK293 cells overexpressing human FOXP3
      Lane 2 : HEK293 cells overexpressing human FOXP3 with Human FOXP3 peptide (ab16809) at 1 µg/ml

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Rabbit Anti-Mouse IgG H&L (HRP) (ab6728) at 1/5000 dilution

      Performed under reducing conditions.

      Predicted band size: 48 kDa


      Exposure time: 45 seconds
    • Immunocytochemistry - Anti-FOXP3 antibody [mAbcam 450] (ab450)
      Immunocytochemistry - Anti-FOXP3 antibody [mAbcam 450] (ab450)This image is courtesy of Alison Banham and Bridget Fox, University of Oxford

      Clone mAbcam 450 used in immunocytochemistry on paraffin embedded transfected COS-1 cytospins.

      1: FOXP1 transfectant
      2: FOXP2 transfectant
      3: FOXP3 transfectant
      4: FOXP4 transfectant

      Strong staining can be seen on the FOXP3 transfectant, with very weak staining on the FOXP1 transfectant and no staining is seen with the other FOXP protein transfectants. This shows that the antibody is largely specific for FOXP3 when used with paraffin embedded cells.

      These images were produced with neat supernatant produced with this clone (the stock of this antibody is purified from supernatant).

    • Immunohistochemistry (Frozen sections) - Anti-FOXP3 antibody [mAbcam 450] (ab450)
      Immunohistochemistry (Frozen sections) - Anti-FOXP3 antibody [mAbcam 450] (ab450)

      IHC image of FOXP3 staining in a section of formalin-fixed frozen normal human tonsil* performed on a Leica BONDTM system using the standard protocol F. The section was then incubated with ab450, 5ug/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

      For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

      *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

    • Immunocytochemistry - Anti-FOXP3 antibody [mAbcam 450] (ab450)
      Immunocytochemistry - Anti-FOXP3 antibody [mAbcam 450] (ab450)This image is courtesy of Alison Banham and Bridget Fox, University of Oxford

      Clone mAbcam 450 used in immunocytochemistry on transfected COS-1 cytospins fixed with acetone for 10 mins.

      1: FOXP1 transfectant
      2: FOXP2 transfectant
      3: FOXP3 transfectant
      4: FOXP4 transfectant

      Staining can be seen on the FOXP3 transfectant, but no staining is seen with the other FOXP protein transfectants, confirming the specificity of this antibody in immunocytochemistry.

      These images were produced with neat supernatant produced with this clone (the stock of this antibody is purified from supernatant).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXP3 antibody [mAbcam 450] (ab450)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXP3 antibody [mAbcam 450] (ab450)

      IHC image of FOXP3 staining in a section of formalin-fixed paraffin-embedded normal human tonsil* performed on a Leica BONDTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab450, 1ug/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

      For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

      *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

    References

    This product has been referenced in:

    • Mu L  et al. The IDH1 Mutation-Induced Oncometabolite, 2-Hydroxyglutarate, May Affect DNA Methylation and Expression of PD-L1 in Gliomas. Front Mol Neurosci 11:82 (2018). Read more (PubMed: 29643764) »
    • Jiang H  et al. Abnormal acetylation of FOXP3 regulated by SIRT-1 induces Treg functional deficiency in patients with abdominal aortic aneurysms. Atherosclerosis 271:182-192 (2018). IP . Read more (PubMed: 29524861) »
    See all 17 Publications for this product

    Publishing research using ab450? Please let us know so that we can cite the reference in this datasheet.

    Customer reviews and Q&As

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    1-2 of 2 Abreviews or Q&A

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-FOXP3 antibody [mAbcam 450]

     Excellent
    Abreviews
    abreview image
    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: Dako Target Retreival Solution
    Permeabilization
    No
    Blocking step
    5 minutes of peroxidase block then 10 minutes of protein block. These are ready-to-use reagents purchased from Dako as blocking agent for 15 minute(s) · Concentration: 100% · Temperature: 20°C
    Read More

    Mr. Antibody Solutions

    Verified customer

    Submitted Apr 07 2009

    Immunoprecipitation abreview for Anti-FOXP3 antibody [mAbcam 450]

     Good
    Abreviews
    Application
    Immunoprecipitation
    Sample
    Human Cell lysate - whole cell (Human FOXP3 transfected COS cells)
    Specification
    Human FOXP3 transfected COS cells
    Immuno-precipitation step
    Protein G
    Read More

    Abcam user community

    Verified customer

    Submitted Jan 26 2006

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