Name: Anti-FOXP3 antibody [EPR22102-37]
Brand: RabMAb
SKU: ab215206
Rating: 5 (4 reviews)

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Western blot - 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

Carrier Free

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

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...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR22102-37
Isotype

IgG
Research areas

Alternative Versions
- Anti-FOXP3 antibody [EPR22102-37] - BSA and Azide free (ab244242)
Compatible Secondaries
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Recombinant Protein
- Recombinant Human FOXP3 protein (His tag) (ab226445)
Related Products
- Universal HIER antigen retrieval reagent (10X) (ab208572)

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
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.
Flow Cyt		1/40.
IHC-P	(4)	1/250. 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.

Notes
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.
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. Recommend to incubate primary antibody at 4C overnight, and use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) as the detection kit.

Application notes

Is unsuitable for ICC/IF or IHC-Fr.

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.
Target information above from: UniProt accession Q9BZS1 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
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
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

Western blot - Anti-FOXP3 antibody [EPR22102-37] (ab215206)

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 seconds

Blocking buffer and concentration: 5% NFDM /TBST
Flow Cytometry (Intracellular) - Anti-FOXP3 antibody [EPR22102-37] (ab215206)

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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXP3 antibody [EPR22102-37] (ab215206)

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).
Immunoprecipitation - Anti-FOXP3 antibody [EPR22102-37] (ab215206)

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).
Western blot - Anti-FOXP3 antibody [EPR22102-37] (ab215206)

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.
Flow Cytometry (Intracellular) - Anti-FOXP3 antibody [EPR22102-37] (ab215206)

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).
Flow Cytometry (Intracellular) - Anti-FOXP3 antibody [EPR22102-37] (ab215206)

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.
Flow Cytometry (Intracellular) - Anti-FOXP3 antibody [EPR22102-37] (ab215206)

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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXP3 antibody [EPR22102-37] (ab215206)

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).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXP3 antibody [EPR22102-37] (ab215206)

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).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXP3 antibody [EPR22102-37] (ab215206)

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).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXP3 antibody [EPR22102-37] (ab215206)

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

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

Datasheet download

Download

Publishing research using ab215206? Please let us know so that we can cite the reference in this datasheet.

ab215206 has been referenced in 3 publications.

Zhu J et al. Interleukin-35 promotes progression of prostate cancer and inhibits anti-tumour immunity. Cancer Cell Int 20:487 (2020). PubMed: 33041668
Lin B et al. Honeysuckle extract relieves ovalbumin-induced allergic rhinitis by inhibiting AR-induced inflammation and autoimmunity. Biosci Rep 39:N/A (2019). PubMed: 31308153
Lv M et al. MicroRNA-155 inhibits the proliferation of CD8+ T cells via upregulating regulatory T cells in vitiligo. Mol Med Rep 20:3617-3624 (2019). PubMed: 31485649

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1-4 of 4 Abreviews or Q&A

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Sample

Human Tissue sections (palatine tonsil)

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: Tris/EDTA

Permeabilization

Specification

palatine tonsil

Blocking step

Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C

Fixative

Paraformaldehyde

Abcam user community

Verified customer

Submitted Aug 05 2020

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Sample

Mouse Tissue sections (Spleen)

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: Tris/EDTA

Permeabilization

Specification

Spleen

Blocking step

Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Fixative

Paraformaldehyde

Mr. Hannes Kaddatz

Verified customer

Submitted Dec 31 2019

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

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Specificity

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Isotype control

Recombinant Protein

Related Products

Applications

The Abpromise guarantee

Target

Function

Involvement in disease

Sequence similarities

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

Datasheet download

Certificate of Compliance

References (3)

Customer reviews and Q&As

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

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

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

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