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RecombinantRabMAb

Recombinant Anti-FOXP3 antibody [EPR22102-37] - BSA and Azide free (ab244242)

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Western blot - Anti-FOXP3 antibody [EPR22102-37] - BSA and Azide free (ab244242)
  • Flow Cytometry (Intracellular) - Anti-FOXP3 antibody [EPR22102-37] - BSA and Azide free (ab244242)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXP3 antibody [EPR22102-37] - BSA and Azide free (ab244242)
  • Immunoprecipitation - Anti-FOXP3 antibody [EPR22102-37] - BSA and Azide free (ab244242)
  • Flow Cytometry (Intracellular) - Anti-FOXP3 antibody [EPR22102-37] - BSA and Azide free (ab244242)
  • Flow Cytometry (Intracellular) - Anti-FOXP3 antibody [EPR22102-37] - BSA and Azide free (ab244242)
  • Flow Cytometry (Intracellular) - Anti-FOXP3 antibody [EPR22102-37] - BSA and Azide free (ab244242)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXP3 antibody [EPR22102-37] - BSA and Azide free (ab244242)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXP3 antibody [EPR22102-37] - BSA and Azide free (ab244242)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXP3 antibody [EPR22102-37] - BSA and Azide free (ab244242)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXP3 antibody [EPR22102-37] - BSA and Azide free (ab244242)
  • Anti-FOXP3 antibody [EPR22102-37] - BSA and Azide free (ab244242)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR22102-37] to FOXP3 - BSA and Azide free
  • Suitable for: IHC-P, WB, IP, Flow Cyt (Intra)
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Unconjugated

Overview

  • Product name

    Anti-FOXP3 antibody [EPR22102-37] - BSA and Azide free
    See all FOXP3 primary antibodies

  • Description

    Rabbit monoclonal [EPR22102-37] to FOXP3 - BSA and Azide free

  • Host species

    Rabbit

  • Specificity

    ab244242 is not recommended for rat in WB.

    ab215206 is  not suitable for endogenous FOXP3 detection in tissues.

  • Tested applications

    Suitable for: IHC-P, WB, IP, Flow Cyt (Intra)more details
    Unsuitable for: ICC/IF or IHC-Fr

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Mouse mammary gland tissue lysate; HEK-293T transfected with FOXP3 (WT) expression vector containing a GFP-myc tag, whole cell lysate.Full-length C-MYC/DDK tagged Recombinant Mouse Foxp3 protein. IHC-P: Human thymus and spleen tissues; Mouse thymus and spleen tissues; Rat thymus tissue. Flow Cyt (intra): 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

    ab244242 is the carrier-free version of ab215206.

    Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

    This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

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

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab244242 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 (1)
1/250 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

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

Recommend to incubate primary antibody at 4C overnight, and use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) as the detection kit.
WB
Use at an assay dependent concentration. Detects a band of approximately 47 kDa (predicted molecular weight: 47 kDa).

ab215206 is  not suitable for endogenous FOXP3 detection in tissues.
IP
Use at an assay dependent concentration.
Flow Cyt (Intra)
Use at an assay dependent concentration.
Notes
IHC-P
1/250 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

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

Recommend to incubate primary antibody at 4C overnight, and use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) as the detection kit.
WB
Use at an assay dependent concentration. Detects a band of approximately 47 kDa (predicted molecular weight: 47 kDa).

ab215206 is  not suitable for endogenous FOXP3 detection in tissues.
IP
Use at an assay dependent concentration.
Flow Cyt (Intra)
Use at an assay dependent concentration.
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

    • Western blot - Anti-FOXP3 antibody [EPR22102-37] - BSA and Azide free (ab244242)
      Western blot - Anti-FOXP3 antibody [EPR22102-37] - BSA and Azide free (ab244242)
      Lane 1 : Anti-FOXP3 antibody [EPR22102-37] (ab215206) at 1/200 dilution
      Lane 2 : Anti-DDDDK tag (Binds to FLAG® tag sequence) antibody [EPR20018-251] (ab205606) at 1/1000 dilution

      All lanes : Full-length C-MYC/DDK tagged Recombinant Mouse Foxp3 protein 10ng

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

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


      Exposure time: 180 seconds


      Blocking buffer and concentration: 5% NFDM /TBST

    • Flow Cytometry (Intracellular) - Anti-FOXP3 antibody [EPR22102-37] - BSA and Azide free (ab244242)
      Flow Cytometry (Intracellular) - Anti-FOXP3 antibody [EPR22102-37] - BSA and Azide free (ab244242)

      Intracellular flow cytometric analysis of4% 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. 

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab215206). 

       

       

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXP3 antibody [EPR22102-37] - BSA and Azide free (ab244242)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXP3 antibody [EPR22102-37] - BSA and Azide free (ab244242)

      Immunohistochemical analysis of paraffin-embedded human thymus tissue labeling FOXP3 with ab215206 at 1/250 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).

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab215206).

    • Immunoprecipitation - Anti-FOXP3 antibody [EPR22102-37] - BSA and Azide free (ab244242)
      Immunoprecipitation - Anti-FOXP3 antibody [EPR22102-37] - BSA and Azide free (ab244242)

      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).

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab215206).

    • Flow Cytometry (Intracellular) - Anti-FOXP3 antibody [EPR22102-37] - BSA and Azide free (ab244242)
      Flow Cytometry (Intracellular) - Anti-FOXP3 antibody [EPR22102-37] - BSA and Azide free (ab244242)

      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). 

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab215206). 

       

       

    • Flow Cytometry (Intracellular) - Anti-FOXP3 antibody [EPR22102-37] - BSA and Azide free (ab244242)
      Flow Cytometry (Intracellular) - Anti-FOXP3 antibody [EPR22102-37] - BSA and Azide free (ab244242)

      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. 

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab215206). 

       

       

    • Flow Cytometry (Intracellular) - Anti-FOXP3 antibody [EPR22102-37] - BSA and Azide free (ab244242)
      Flow Cytometry (Intracellular) - Anti-FOXP3 antibody [EPR22102-37] - BSA and Azide free (ab244242)

      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. 

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab215206). 

       

       

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXP3 antibody [EPR22102-37] - BSA and Azide free (ab244242)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXP3 antibody [EPR22102-37] - BSA and Azide free (ab244242)

      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).

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab215206).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXP3 antibody [EPR22102-37] - BSA and Azide free (ab244242)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXP3 antibody [EPR22102-37] - BSA and Azide free (ab244242)

      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).

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab215206).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXP3 antibody [EPR22102-37] - BSA and Azide free (ab244242)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXP3 antibody [EPR22102-37] - BSA and Azide free (ab244242)

      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).

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab215206).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXP3 antibody [EPR22102-37] - BSA and Azide free (ab244242)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXP3 antibody [EPR22102-37] - BSA and Azide free (ab244242)

      Immunohistochemical analysis of paraffin-embedded human spleen tissue labeling FOXP3 with ab215206 at 1/500 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).

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab215206).

    • Anti-FOXP3 antibody [EPR22102-37] - BSA and Azide free (ab244242)
      Anti-FOXP3 antibody [EPR22102-37] - BSA and Azide free (ab244242)

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    References (0)

    Publishing research using ab244242? Please let us know so that we can cite the reference in this datasheet.

    ab244242 has not yet been referenced specifically in any publications.

    Customer reviews and Q&As

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    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-FOXP3 antibody [EPR22102-37] - BSA and Azide free

     Good
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (Spleen)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Tris-EDTA pH 9
    Permeabilization
    Yes - Tween, Triton
    Specification
    Spleen
    Blocking step
    1% BSA, 2% donkey serum, 0.1% gelatin as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
    Fixative
    Paraformaldehyde
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Aug 02 2021

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