Recombinant
RabMAb

Recombinant Anti-FOXP3 antibody [EPR22102-37] (ab215206)

Overview

  • Product name

    Anti-FOXP3 antibody [EPR22102-37]
    See all FOXP3 primary antibodies
  • Description

    Rabbit monoclonal [EPR22102-37] to FOXP3
  • Host species

    Rabbit
  • Specificity

    ab215206 is not recommended for rat in WB.

  • Tested applications

    Suitable for: WB, IP, Flow Cyt, IHC-Pmore details
    Unsuitable for: ICC/IF or IHC-Fr
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment within Human FOXP3 aa 50 to the C-terminus. The exact sequence is proprietary.
    Database link: Q9BZS1

  • Positive control

    • WB: Human tonsil, mammary gland and breast cancer tissue lysates; Mouse mammary gland tissue lysate; HEK-293T transfected with FOXP3 (WT) expression vector containing a GFP-myc tag, whole cell lysate. IHC-P: Human thymus and spleen tissues; Mouse thymus and spleen tissues; Rat thymus tissue. Flow Cyt: HEK-293T cell line transfected with a GFP-tagged FOXP3 expression construct. IP: HEK-293T cells transfected with a GFP-tagged FOXP3 expression construct whole cell lysate.
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab215206 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
WB 1/1000. Detects a band of approximately 47 kDa (predicted molecular weight: 47 kDa).
IP 1/30.
Flow Cyt 1/40.
IHC-P 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

Use at 1/1000 dilution for rat and mouse tissues.

  • Application notes
    Is unsuitable for ICC/IF or IHC-Fr.
  • 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

    • Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) cell line transfected with a GFP-tagged FOXP3 expression construct labeling FOXP3 with ab215206 at 1/40 dilution (Right) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (Left). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) (ab150079), at 1/2000 dilution was used as the secondary antibody.

    • Immunohistochemical analysis of paraffin-embedded human thymus tissue labeling FOXP3 with ab215206 at 1/1000 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on regulatory T-cells in human thymus (PMID: 16380964) is observed. Counter stained with hematoxylin.
      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

      Heat mediated antigen retrieval using ab208572 (Universal HIER antigen retrieval reagent).

    • FOXP3 was immunoprecipitated from 0.35 mg of HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) cells transfected with a GFP-tagged FOXP3 expression construct whole cell lysate with ab215206 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab215206 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.

      Lane 1: HEK-293T cells transfected with a GFP-tagged FOXP3 expression construct whole cell lysate 10 μg (Input).
      Lane 2: ab215206 IP in HEK-293T cells transfected with a GFP-tagged FOXP3 expression construct whole cell lysate (+). 
      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab215206 in HEK-293T cells transfected with a GFP-tagged FOXP3 expression construct whole cell lysate (-).

      Blocking and dilution buffer and concentration: 5% NFDM/TBST.
      Exposure time: 3 seconds.

      FOXP3 isoforms have been described in the literature. (PMID: 19568423).

    • All lanes : Anti-FOXP3 antibody [EPR22102-37] (ab215206) at 1/1000 dilution

      Lane 1 : HEK-293T (human embryonic kidney) transfected with an empty vector (vector control), containing a GFP-myc tag, whole cell lysate
      Lane 2 : HEK-293T transfected with FOXP3 (WT) expression vector containing a GFP-myc tag, whole cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Predicted band size: 47 kDa
      Observed band size: 73 kDa why is the actual band size different from the predicted?


      Exposure time: 6 seconds


      Blocking and dilution buffer: 5% NFDM/TBST.

    • All lanes : Anti-FOXP3 antibody [EPR22102-37] (ab215206) at 1/1000 dilution

      Lane 1 : Human tonsil tissue lysate
      Lane 2 : Human mammary gland tissue lysate
      Lane 3 : Human breast cancer tissue lysate
      Lane 4 : Mouse mammary gland tissue lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution

      Predicted band size: 47 kDa
      Observed band size: 47 kDa



      Exposure time : Lanes 1 & 2: 26 seconds. Lanes 3 & 4: 3 minutes.

      Blocking and dilution buffer: 5% NFDM/TBST.

    • Human PBMCs were stained with Alexa Fluor® 647 conjugated anti-human CD4 and BV421 conjugated anti-human CD25. Cells were then fixed with 2% PFA for 10min and permeabilized with True-Nuclear™ permeabilization buffer, followed by intracellular staining with rabbit IgG (ab172730, Left) or anti-FOXP3 RabMab (Right, 1/40). ab98462, Dylight® 488-conjugated goat anti-rabbit IgG was used as the secondary at a dilution of 1/2000.

      Gating strategy and expression pattern is consistent with literature (PMID: 27330808).

    • Mouse splenocytes were stained with Alexa Fluor® 647 conjugated anti-mouse CD4. Cells were then fixed with 2% PFA for 10min and permeabilized with True-Nuclear™ permeabilisation buffer, followed by intracellular staining with rabbit IgG (ab172730, Left) or anti-FOXP3 RabMab (Right, 1/40). ab150077, Alexa Fluor® 488-conjugated goat anti-rabbit IgG was used as the secondary at a dilution of 1/2000.

    • Rat splenocytes were stained with Alexa Fluor® 647 conjugated anti-rat CD4. Cells were then fixed with 2% PFA for 10min and permeabilized with True-Nuclear™ permeabilization buffer, followed by intracellular staining with rabbit IgG (ab172730, Left) or anti-FOXP3 RabMab (Right, 1/40). ab150077, Alexa Fluor® 488-conjugated goat anti-rabbit IgG was used as the secondary at a dilution of 1/2000.

    • Immunohistochemical analysis of paraffin-embedded rat thymus tissue labeling FOXP3 with ab215206 at 1/1000 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on regulatory T-cells in rat thymus (PMID: 16380964) is observed. Counter stained with hematoxylin.
      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

      Heat mediated antigen retrieval using ab208572 (Universal HIER antigen retrieval reagent).

    • Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling FOXP3 with ab215206 at 1/1000 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on regulatory T-cells in mouse spleen (PMID: 16380964) is observed. Counter stained with hematoxylin.
      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

      Heat mediated antigen retrieval using ab208572 (Universal HIER antigen retrieval reagent).

    • Immunohistochemical analysis of paraffin-embedded mouse thymus tissue labeling FOXP3 with ab215206 at 1/1000 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on regulatory T-cells in mouse thymus (PMID: 16380964) is observed. Counter stained with hematoxylin.
      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

      Heat mediated antigen retrieval using ab208572 (Universal HIER antigen retrieval reagent).

    • Immunohistochemical analysis of paraffin-embedded human spleen tissue labeling FOXP3 with ab215206 at 1/1000 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on regulatory T-cells in human spleen (PMID: 16380964) is observed. Counter stained with hematoxylin.
      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

      Heat mediated antigen retrieval using ab208572 (Universal HIER antigen retrieval reagent).

    References

    ab215206 has not yet been referenced specifically in any publications.

    Customer reviews and Q&As

    1-2 of 2 Abreviews or Q&A

    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (Lymph node)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Tris/EDTA
    Permeabilization
    Yes - H2O2
    Specification
    Lymph node
    Fixative
    Formaldehyde

    Abcam user community

    Verified customer

    Submitted Aug 19 2019

    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (Tumor)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Leica ER2 solution
    Specification
    Tumor
    Fixative
    Formaldehyde

    Zohar Gavish

    Verified customer

    Submitted May 13 2019

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