Astana Biomed Group, an authorized Abcam distributor in Central Asia

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: Flow Cyt, IHC-P, WB, IP
  • Reacts with: Mouse, Rat, Human

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: Flow Cyt, IHC-P, WB, IPmore 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: 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. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with

    ab244242 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    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.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

Properties

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

    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.

    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 1/1000 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).

    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 - Anti-FOXP3 antibody [EPR22102-37] - BSA and Azide free (ab244242)
    Flow Cytometry - 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 - Anti-FOXP3 antibody [EPR22102-37] - BSA and Azide free (ab244242)
    Flow Cytometry - 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 - Anti-FOXP3 antibody [EPR22102-37] - BSA and Azide free (ab244242)
    Flow Cytometry - 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 1/1000 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).

    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)

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