Anti-FOXP3 antibody [EPR22102-37] (ab215206)
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
- Suitable for: WB, IP, Flow Cyt, IHC-P
- Reacts with: Mouse, Rat, Human
Overview
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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.
According to our preliminary WB data, this antibody failed to detect endogenous expression of FOXP3 in the tissues tested in WB, such as thymus. It might work in CD4+CD25+ Treg cells with abundant FOXP3, but we don’t have experimental data to support that.
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Tested Applications & Species
See all applications and species dataApplication Species Flow Cyt MouseRatHumanIHC-P MouseRatHumanIP HumanWB MouseHuman -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: 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.
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General notes
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
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
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR22102-37 -
Isotype
IgG -
Research areas
Images
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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 (Anti-DDDDK tag, 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 secondsBlocking buffer and concentration: 5% NFDM /TBST
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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.
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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).
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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).
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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 secondsBlocking and dilution buffer: 5% NFDM/TBST.
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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).
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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.
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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.
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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).
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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).
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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).
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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).
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