Recombinant 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, IHC-P, Flow Cyt (Intra), mIHC
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
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
The rat recommendation is based on the IHC results. We do not guarantee WB for rat.
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
Suitable for: WB, IP, IHC-P, Flow Cyt (Intra), mIHCmore 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.
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Positive control
- WB: HEK-293T transfected with FOXP3 (WT) expression vector with a GFP-myc tag, whole cell lysate. Full-length C-MYC/DDK tagged Recombinant Mouse Foxp3 protein. IHC-P: Human, mouse, rat thymus tissues, human and mouse spleen tissues, human tonsil tissue. Flow Cyt (intra): HEK-293T transfected with a GFP-tagged FOXP3 expression construct. IP: HEK-293T 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
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
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 |
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WB |
1/1000. Detects a band of approximately 47 kDa (predicted molecular weight: 47 kDa).
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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IP |
1/30.
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IHC-P | (5) |
1/100 - 1/1000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Please adjust the dilution factor according to tissues from different species. Recommend performing IHC staining on a Leica Biosystems BOND® RX instrument because BOND® RX instrument assay is stronger than overnight incubation assay. |
Flow Cyt (Intra) |
1/40.
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mIHC |
1/100.
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Notes |
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WB
1/1000. Detects a band of approximately 47 kDa (predicted molecular weight: 47 kDa). 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. |
IP
1/30. |
IHC-P
1/100 - 1/1000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. Please adjust the dilution factor according to tissues from different species. Recommend performing IHC staining on a Leica Biosystems BOND® RX instrument because BOND® RX instrument assay is stronger than overnight incubation assay. |
Flow Cyt (Intra)
1/40. |
mIHC
1/100. |
Target
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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
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Database links
- Entrez Gene: 50943 Human
- Entrez Gene: 20371 Mouse
- Entrez Gene: 317382 Rat
- Omim: 300292 Human
- SwissProt: Q9BZS1 Human
- SwissProt: Q99JB6 Mouse
- Unigene: 247700 Human
- Unigene: 182291 Mouse
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Alternative names
- AIID antibody
- DIETER antibody
- Forkhead box P3 antibody
see all
Images
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Immunohistochemical analysis of paraffin-embedded human tonsil tissue labelling FOXP3 with ab215206 at 1:100 dilution (3.67 μg/ml).
Image A: LeicaDS9800 (Bond™ Polymer Refine Detection) secondary antibody used at a ready to use concentration. Nuclear staining on human tonsil. The section was incubated with ab215206 for 30min at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) for 20 mins.
Image B: Goat Anti-Rabbit IgG H&L (HRP polymer) secondary antibody used at a ready to use concentration. Nuclear staining on human tonsil. The section was incubated with ab215206 overnight at +4 ℃. Counterstained with Hematoxylin.
Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).
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10-color fluorescence multiplex immunohistochemical analysis of human lung cancer tissue (formalin-fixed paraffin-embedded section).
Merged staining of anti-FOXP3 (ab215206; Cyan; TG540N), anti-PD1 (ab52587; Red; TG700N), anti-CD163 (ab182422; Brown; TG650N), anti-HLA-DR (ab92511; Yellow; TG570N), anti-CD4 (ab133616; Violet; TG620N), anti-CD8 alpha (ab101500; Purple; TG540S), anti-CD20 (ab9475; Grey; TG660S), anti-CD68 (ab192847; Green; TG520N), anti-Cytokeratin 19 (ab52625; Light blue; TG440N). TG470SN (dark blue) was used as a nuclear counter stain. The inset image shows the separate FOXP3 signal.
The section was incubated in nine rounds of staining; in the order of ab215206 (1/100 dilution), ab52587 (1/200 dilution), ab182422 (1/300 dilution), ab92511 (1/200 dilution), ab133616 (1/600 dilution), ab101500 (1/300 dilution), ab9475 (1/100 dilution), ab192847 (1/300 dilution), ab52625 (1/400 dilution); each using a separate fluorescent tyramide signal amplification system.
Sodium citrate antigen retrieval (pH6.0) was used in between rounds of tyramide signal amplification to remove the antibody from the previous round, to avoid any cross-reactivity.
Image acquisition was performed with TissueFAXS Spectra (TissueGnostics).
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10-color fluorescence multiplex immunohistochemical analysis of human lung cancer tissue (formalin-fixed paraffin-embedded section).
Merged staining of anti-FOXP3 (ab215206; Cyan; TG540N), anti-PD1 (ab52587; Violet; TG700N), anti-CD163 (ab182422; Red; TG650N), anti-HLA-DR (ab92511; Yellow; TG570N), anti-CD4 (ab133616; Orange; TG620N), anti-CD8 alpha (ab101500; Purple; TG540S), anti-CD20 (ab9475; Grey; TG660S), anti-CD68 (ab192847; Green; TG520N), anti-Cytokeratin 19 (ab52625; Light blue; TG440N). TG470SN (dark blue) was used as a nuclear counter stain.
The section was incubated in nine rounds of staining; in the order of ab215206 (1/100 dilution), ab52587 (1/200 dilution), ab182422 (1/300 dilution), ab92511 (1/200 dilution), ab133616 (1/600 dilution), ab101500 (1/300 dilution), ab9475 (1/100 dilution), ab192847 (1/300 dilution), ab52625 (1/400 dilution); each using a separate fluorescent tyramide signal amplification system.
Sodium citrate antigen retrieval (pH6.0) was used in between rounds of tyramide signal amplification to remove the antibody from the previous round, to avoid any cross-reactivity.
Image acquisition was performed with TissueFAXS Spectra (TissueGnostics).
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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.
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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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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).
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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 1/500 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).
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.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (45)
ab215206 has been referenced in 45 publications.
- Zheng Y et al. Radiation combined with KRAS-MEK inhibitors enhances anticancer immunity in KRAS-mutated tumor models. Transl Res 252:79-90 (2023). PubMed: 35948200
- Bertho A et al. Evaluation of the Role of the Immune System Response After Minibeam Radiation Therapy. Int J Radiat Oncol Biol Phys 115:426-439 (2023). PubMed: 35985455
- Yang Y et al. Targeting the STAT5A/IDO1 axis overcomes radioresistance and reverses the immunosuppressive tumor microenvironment in NSCLC. Int J Oncol 62:N/A (2023). PubMed: 36453241
- Song G et al. Single-cell transcriptomic analysis suggests two molecularly subtypes of intrahepatic cholangiocarcinoma. Nat Commun 13:1642 (2022). PubMed: 35347134
- Wang T et al. Regulation of Th17/Treg Balance by 27-Hydroxycholesterol and 24S-Hydroxycholesterol Correlates with Learning and Memory Ability in Mice. Int J Mol Sci 23:N/A (2022). PubMed: 35457188