Overview

  • Product nameAnti-FOXP3 antibody [236A/E7]
    See all FOXP3 primary antibodies
  • Description
    Mouse monoclonal [236A/E7] to FOXP3
  • Tested applicationsSuitable for: IHC-P, IHC-Fr, WB, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Human, Cynomolgus Monkey, Rhesus monkey
  • Immunogen

    FOXP3 fusion protein.

  • Positive control
    • Human tonsil, HEK293T cell lysate overexpressing Human FOXP3

Properties

Applications

Our Abpromise guarantee covers the use of ab20034 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-P Use a concentration of 10 µg/ml. Perform heat mediated antigen retrieval via the microwave method before commencing with IHC staining protocol.
IHC-Fr Use a concentration of 10 µg/ml.
WB Use a concentration of 4 - 5 µg/ml. Detects a band of approximately 50 kDa (predicted molecular weight: 47 kDa).
ICC/IF Use at an assay dependent concentration.

Target

  • FunctionProbable transcription factor. Plays a critical role in the control of immune response.
  • Involvement in diseaseDefects 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 similaritiesContains 1 C2H2-type zinc finger.
    Contains 1 fork-head DNA-binding domain.
  • Cellular localizationNucleus.
  • 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

Anti-FOXP3 antibody [236A/E7] images

  • All lanes : Anti-FOXP3 antibody [236A/E7] (ab20034) at 5 µg/ml

    Lane 1 : Human thymus
    Lane 2 : Mouse thymus
    Lane 3 : Rat thymus

    Lysates/proteins at 20 µg per lane.

    Secondary
    IR-labelled goat anti-mouse (green; 800) at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size : 47 kDa
    Observed band size : 26,50 kDa (why is the actual band size different from the predicted?)

    This blot was produced using a 4-12% Bis-Tris gel under the MOPS buffer system. The gel was run at 200V for 60 minutes before being transferred onto a nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour before being incubated with Anti-FOXP3 antibody [236A/E7] (ab20034; 5 microgram per mL), overnight at 4°C. Antibody binding was detected using infrared labelled goat anti-mouse (green; 1:10000) for 1 hour at room temperature before imaging.

  • IHC image of Foxp3 staining in Human normal tonsil formalin fixed paraffin embedded tissue section*, performed on a Leica Bond™ 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 20 mins. The section was then incubated with ab20034, 5µg/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

  • All lanes : Anti-FOXP3 antibody [236A/E7] (ab20034) at 5 µg/ml

    Lane 1 : HEK293T cell lysate
    Lane 2 : HEk293T cell lysate overexpressing Human FOXP3
    Lane 3 : Human tonsil tissue lysate

    Lysates/proteins at 20 µg per lane.


    Performed under reducing conditions.

    Predicted band size : 47 kDa

    ab20034 detects Human FOXP3 protein at ~50 kDa in HEK293T cells overexpressing the protein. It also detects FOXP3 in Human tonsil tissue lysate, however this band is significantly weaker in endogenous conditions. Upon higher exposure, weak bands can also be observed in HEK293T cell lysate.

    This blot was produced using a 4-12% Bis-Tris gel under the MOPS buffer system. The gel was run at 200V for 60 minutes before being transferred onto a nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour before being incubated with Anti-FOXP3 antibody [236A/E7] (ab20034; 5 microgram per mL) overnight at 4°C. Antibody binding was detected using infrared labelled goat anti-mouse (green; 1:10000) for 1 hour at room temperature before imaging.

  • ab20034 staining FOXP3 in Cynomolgus Monkey Spleen tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 10% donkey serum for 20 minutes at room temperature; antigen retrieval was by heat mediation using EDTA, pH9.0. Samples were incubated with primary antibody (1/100) for 30 minutes. A Biotin-conjugated Donkey anti-mouse polyclonal (1/2000) was used as the secondary antibody.

    See Abreview

  • ab20034 staining FOXP3 in Human breast cancer tissue sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with acetone/methanol (1:2), permeabilized with TBS-TritonX-100 and blocked with 10% serum for 10 minutes at 20°C. Samples were incubated with primary antibody (1/50 in PBT (PBS/BSA/Tween)) for 30 minutes at 20°C. An undiluted HRP-conjugated rat anti-mouse/rabbit IgG polyclonal was used as the secondary antibody.

    See Abreview

  • ab20034 used on paraffin embedded human tonsil sections.
    Double staining: CD30 in red, FOXP3 in black, blue is haemotoxylin counterstaining of nuclei.

    As expected FOXP3 stains a sub-population of T cells.

  • ab20034 used on paraffin embedded human tonsil sections.
    FOXP3 staining is black, blue is haemotoxylin counterstaining of nuclei.

    As expected FOXP3 stains a sub-population of T cells.

  • ab20034 staining FOXP3 in human colon tissue section by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue underwent formaldehyde fixation before heat mediated antigen retrieval and then blocking with 10% serum for 2 hours at 21°C. The primary antibody was diluted 1/50 and incubated with sample for 2 hours at 21°C. An Alexa Fluor®488-conjugated rabbit polyclonal to mouse IgG was used undiluted as secondary antibody.

    See Abreview

References for Anti-FOXP3 antibody [236A/E7] (ab20034)

This product has been referenced in:
  • Nguyen N  et al. Tumor infiltrating lymphocytes and survival in patients with head and neck squamous cell carcinoma. Head Neck 38:1074-84 (2016). Human . Read more (PubMed: 26879675) »
  • Gu L  et al. Rapamycin ameliorates CCl4-induced liver fibrosis in mice through reciprocal regulation of the Th17/Treg cell balance. Mol Med Rep 14:1153-61 (2016). WB ; Mouse . Read more (PubMed: 27315465) »

See all 80 Publications for this product

Product Wall

Thank you for your email. I am sorry to hear that you are having problems with this antibody.

I have checked the provided Image and details; I however have few questions regarding the protocol;

- What is the tissue type and species...

Read More
Application Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample Mouse Tissue sections (Spleen)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: 10 mM sodium citrate
Specification Spleen
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Jun 03 2016

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (Liver/Colon/Tonsil)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Ph6 20 mins 100oC
Permeabilization No
Specification Liver/Colon/Tonsil
Blocking step No blocking step used for 10 minute(s) · Concentration: 3% · Temperature: 37°C
Fixative Formaldehyde
Username

Abcam user community

Verified customer

Submitted May 30 2016

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (Skin melanoma)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Low pH
Permeabilization No
Specification Skin melanoma
Blocking step Hydrogen Peroxide as blocking agent for 5 minute(s) · Concentration: 5% · Temperature: RT°C
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Nov 02 2015


For paraffin embedded tissues an initial automated dewaxing and rehydration step followed by heat-induced (100°C for 20 min) or enzyme-induced antigen retrieval (10 to 15 min, Bond Enzyme Pretreatment Kit, Leica Biosystems) was performed.
...

Read More

We do not know the exact location of the epitope that the antibody recognizes but we do know it is within the amino acid range aa107-aa196 of isoform 1 of the protein sequence at UniProt accession Q9BZS1. Here is a link to the UniProt entry.

Read More

Le adjunto en mi mail un esquema que sumariza los pasos de una immunohistoquímica y en la que se detalla el orden a seguir para realizar la tinción. El paso de permeabilización del tejido es anterior a la incubación con el a...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (Human head & neck squamous cell carcinoma)
Specification Human head & neck squamous cell carcinoma
Fixative Paraformaldehyde
Permeabilization Yes - Triton X-100 (0.1%) for 5 min
Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 22°C
Username

Abcam user community

Verified customer

Submitted Jan 28 2013

Thank you for your inquiry.

I am happy to confirm that the immunogen range is from aa107-aa196 of isoform 1 on Swissprot link http://www.uniprot.org/uniprot/Q9BZS1.

We have no additional (experimental) data about the isoform specifi...

Read More

Thank you for your reply.

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.

If there is any concrete...

Read More

1-10 of 44 Abreviews or Q&A

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