Human/Mouse FoxP3 Antibody

R&D Systems | Catalog # MAB8214

Recombinant Monoclonal Antibody.
R&D Systems

Key Product Details

Validated by

Orthogonal Validation, Biological Validation

Species Reactivity

Validated:

Human, Mouse

Cited:

Human, Mouse, Transgenic Mouse

Applications

Validated:

Multiplex Immunofluorescence, Immunohistochemistry, Flow Cytometry, Dual RNAscope ISH-IHC Compatible, Immunocytochemistry, COMET

Cited:

Immunohistochemistry, Immunohistochemistry-Paraffin

Label

Unconjugated

Antibody Source

Recombinant Monoclonal Rabbit IgG Clone # 1054C
View all FoxP3 Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human FoxP3
Met1-Leu71
Accession # Q9BZS1

Specificity

Detects human FoxP3 in direct ELISAs. Detects human and mouse FoxP3 in flow cytometry.

Clonality

Monoclonal

Host

Rabbit

Isotype

IgG

Scientific Data Images for Human/Mouse FoxP3 Antibody

Detection of FOXP3 in Hodgkin's Lymphoma via Multiplex Immunofluorescence staining on COMET™

FOXP3 was detected in immersion fixed paraffin-embedded sections of Hodgkin's Lymphoma using Rabbit Anti-Human FOXP3 Monoclonal Antibody (Catalog # MAB8214) at 20 µg/mL at 37 ° Celsius for 4 minutes. Before incubation with the primary antibody, tissue underwent an all-in-one dewaxing and antigen retrieval preprocessing using PreTreatment Module (PT Module) and Dewax and HIER Buffer H (pH 9). Tissue was stained using the Alexa Fluor™ Plus 647 Goat anti-Rabbit IgG Secondary Antibody at 1:200 at 37 ° Celsius for 2 minutes. (Yellow; Lunaphore Catalog # DR647RB) and counterstained with DAPI (blue; Lunaphore Catalog # DR100). Specific staining was localized to the nucleus. Protocol available in COMET™ Panel Builder.

Detection of FOXP3 in Mouse Thymus via seqIF™ staining on COMET™​

FOXP3 Antibody was detected in immersion fixed paraffin-embedded sections of mouse Thymus using Rabbit Anti-Mouse FOXP3 Monoclonal Antibody (Catalog # MAB8214) at 20ug/mL at 37 ° Celsius for 4 minutes. Before incubation with the primary antibody, tissue underwent an all-in-one dewaxing and antigen retrieval preprocessing using PreTreatment Module (PT Module) and Dewax and HIER Buffer H (pH 9; Epredia Catalog # TA-999-DHBH). Tissue was stained using the Alexa Fluor™ Plus 647 Goat anti-Rabbit IgG Secondary Antibody at 1:200 at 37 ° Celsius for 2 minutes. (Yellow; Lunaphore Catalog # DR647RB) and counterstained with DAPI (blue; Lunaphore Catalog # DR100). Specific staining was localized to the nucleus. Protocol available in COMET™ Panel Builder.

Detection of FoxP3 antibody in Human PBMCs stimulated to induce Tregs antibody by Flow Cytometry.

Detection of FoxP3 in Human PBMCs stimulated to induce Tregs by Flow Cytometry.

Human peripheral blood mononuclear cells (PBMCs) either (A) untreated or (B) stimulated to induce Regulatory T Cells (Tregs) with Recombinant Human TGF-beta 1 (Catalog # 240-B) and Recombinant Human IL-2 (Catalog # 202-IL) for 2 days were stained with Rabbit Anti-Human/Mouse FoxP3 Monoclonal Antibody (Catalog # MAB8214) followed by Phycoerythrin-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # F0110) and Mouse Anti-Human CD4 APC-conjugated Monoclonal Antibody (Catalog # FAB3791A). Quadrant markers were set based on control antibody staining (Catalog # AB-105-C). To facilitate intracellular staining, cells were fixed and permeabilized with FlowX FoxP3 Fixation & Permeabilization Buffer Kit (Catalog # FC012).

Detection of FoxP3 antibody in Mouse Splenocytes antibody by Flow Cytometry.

Detection of FoxP3 in Mouse Splenocytes by Flow Cytometry.

Mouse splenocytes were stained with Rat Anti-Mouse CD4 APC-conjugated Monoclonal Antibody (Catalog # FAB554A) and either (A) Rabbit Anti-Human/Mouse FoxP3 Monoclonal Antibody (Catalog # MAB8214) or (B) Normal Rabbit IgG Control (Catalog # AB-105-C) followed by Phycoerythrin-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # F0110). To facilitate intracellular staining, cells were fixed and permeabilized with FlowX FoxP3 Fixation & Permeabilization Buffer Kit (Catalog # FC012).

FoxP3 antibody in HeLa Human Cell Line by Immunocytochemistry (ICC).

FoxP3 in HeLa Human Cell Line.

FoxP3 was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line using Rabbit Anti-Human/Mouse FoxP3 Monoclonal Antibody (Catalog # MAB8214) at 8 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red, upper panel; Catalog # NL004) and counterstained with DAPI (blue, lower panel). Specific staining was localized to nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

FoxP3 antibody in Human Tonsil by Immunohistochemistry (IHC-P).

FoxP3 in Human Tonsil.

FoxP3 was detected in immersion fixed paraffin-embedded sections of human tonsil using Rabbit Anti-Human/Mouse FoxP3 Monoclonal Antibody (Catalog # MAB8214) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Rabbit HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS005) and counterstained with hematoxylin (blue). Specific staining was localized to nuclei. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

FoxP3 antibody in Human Ovarian Cancer Tissue by Immunohistochemistry (IHC-P).

FoxP3 in Human Ovarian Cancer Tissue.

FoxP3 was detected in immersion fixed paraffin-embedded sections of human ovarian cancer tissue using Rabbit Anti-Human/Mouse FoxP3 Monoclonal Antibody (Catalog # MAB8214) at 5 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Rabbit IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC003). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to nuclei. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

FoxP3 antibody in Human Tonsil Using Dual RNAscope®ISH and IHC.

FoxP3 in Human Tonsil Using Dual RNAscope®ISH and IHC.

FoxP3 mRNA (red) and protein (green) was detected in formalin-fixed paraffin-embedded tissue sections of human tonsil probed with ACD RNAScope®Probe (Catalog # 418471) followed by immunohistochemistry using R&D Systems Rabbit Anti-Human/Mouse FoxP3 Monoclonal Antibody (Catalog# MAB8214) at 5ug/mL for 1 hour at room temperature followed by incubation with the Anti-Rabbit IgG VisUCyte HRP Polymer Antibody (R&D Systems, Catalog # VC003). Tissue was stained using ACD RNAscope®2.5 HD Duplex Detection Reagents (Catalog # 322500).

Detection of Human FoxP3 by Immunocytochemistry/Immunofluorescence

Detection of Human FoxP3 by Immunocytochemistry/Immunofluorescence

Multiplex fluorescent immunohistochemistry staining. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31192136), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human FoxP3 by Immunocytochemistry/Immunofluorescence

Detection of Human FoxP3 by Immunocytochemistry/Immunofluorescence

Multiplex fluorescent immunohistochemistry staining. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31192136), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of FoxP3 in Mouse Thymus.

FoxP3 was detected in immersion fixed paraffin-embedded sections of mouse thymus using Rabbit Anti-Human/Mouse FoxP3 Monoclonal Antibody (Catalog # MAB8214) at 15 µg/ml for 1 hour at room temperature followed by incubation with the Anti-Rabbit IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC003). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using VisUCyte Antigen Retrieval Reagent-Basic (Catalog # VCTS021). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to the nucleus. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Applications for Human/Mouse FoxP3 Antibody

Application
Recommended Usage

COMET

Optimal dilutions of this antibody should be experimentally determined.

Dual RNAscope ISH-IHC Compatible

5-25 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human tonsil

Flow Cytometry

0.25 µg/106 cells
Sample: Human peripheral blood mononuclear cells (PBMCs) stimulated to induce Regulatory T Cells (Tregs) with Recombinant Human TGF‑ beta 1 (Catalog # 240-B) and Recombinant Human IL‑2 (Catalog # 202-IL) and mouse splenocytes were fixed and permeabilized with FlowX FoxP3 Fixation & Permeabilization Buffer Kit (Catalog # FC012)

Immunocytochemistry

8-25 µg/mL
Sample: Immersion fixed HeLa human cervical epithelial carcinoma cell line

Immunohistochemistry

5-25 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human tonsil, human ovarian cancer tissue, and mouse thymus tissue

Multiplex Immunofluorescence

20 µg/mL
Sample: Immersion fixed paraffin-embedded sections of Hodgkin's Lymphoma and Mouse Thymus

Reviewed Applications

Read 2 reviews rated 4 using MAB8214 in the following applications:

Flow Cytometry Panel Builder

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Advanced Features

  • Spectra Viewer - Custom analysis of spectra from multiple fluorochromes
  • Spillover Popups - Visualize the spectra of individual fluorochromes
  • Antigen Density Selector - Match fluorochrome brightness with antigen density
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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Protein A or G purified from cell culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. See Certificate of Analysis for details.
*Small pack size (-SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: FoxP3

Human FoxP3 is a 47 kDa member of the P subclass of the FOX (forkhead box) family of transcription factors. It contains a Leu-rich repeat, a C2H2 zinc finger region, and a C-terminal FKH (fork head), DNA-binding domain. Three isoforms for FoxP3 have been reported. All three isoforms share the sequence used as the immunogen. FoxP3 directly associates with NFAT and NFkB, suppressing their activity in CD4+ T cells. In human, FoxP3 is found in CD4+, CD8+ and CD4+CD25+ T cells. Over the region used for immunization of the amino acid sequence, mouse FoxP3 is 83% to 88% identical to rat, human, canine, and bovine FoxP3.

Long Name

Forkhead Box P3

Alternate Names

AIID, DIETER, IPEX, JM2, PIDX, SCURFIN, XPID

Entrez Gene IDs

50943 (Human); 20371 (Mouse)

Gene Symbol

FOXP3

UniProt

Q9BZS1

Additional FoxP3 Products

Product Documents for Human/Mouse FoxP3 Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

Download SDS

Product Specific Notices for Human/Mouse FoxP3 Antibody

For research use only

Citations for Human/Mouse FoxP3 Antibody

Customer Reviews for Human/Mouse FoxP3 Antibody (2)

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Customer Images

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  • Human/Mouse FoxP3 Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: MCF-7 human breast cancer cell line
    Species: Human
    Verified Customer | Posted 05/25/2017
  • Human/Mouse FoxP3 Antibody
    Name: Manuel Flores
    Application: Immunohistochemistry-Paraffin
    Sample Tested: FFPE Tonsil and FFPE human tonsil
    Species: Human
    Verified Customer | Posted 03/03/2017
    Simultaneous labelling of FoxP3 (magenta) and CD4 (green) on FFPE human tonsil sections. Nuclei stained with DAPI (blue). FoxP3 Antibody (1054C) MAB8214. Secondary Ab used is donkey anti-rabbit A647. Arrows pointing at FoxP3+ cells.
    Human/Mouse FoxP3 Antibody MAB8214

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Protocols

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

FAQs for Human/Mouse FoxP3 Antibody

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  • Q: What detection reagent was used for IHC, to create the Dual RNAscope ISH-IHC image on the datasheet for Catalog # MAB8214? 

    A: Green Chromogen from ACD (ACD Catalog # 322550) was ued for IHC detection. See ACD website (https://acdbio.com/store/catalog/product/view/id/22/) for protocol and handling recommendations.  Extensive washing with buffers containing PBS is discouraged as it may cause decolorization. It is critical that after staining with green chromogen, slides with tissues are allowed to dry at room temperature prior to using a permanent mounting media.

  • Q: What is the immunogen sequence of this FoxP3 antibody?

    A: A sequence from Met1 - Leu71.

  • Q: What detection reagent was used for IHC, to create the Dual RNAscope ISH-IHC image on the datasheet for Catalog # MAB8214? 

    A: Green Chromogen from ACD (ACD Catalog # 322550) was ued for IHC detection. See ACD website (https://acdbio.com/store/catalog/product/view/id/22/) for protocol and handling recommendations.  Extensive washing with buffers containing PBS is discouraged as it may cause decolorization. It is critical that after staining with green chromogen, slides with tissues are allowed to dry at room temperature prior to using a permanent mounting media.

  • Q: What is the immunogen sequence of this FoxP3 antibody?

    A: A sequence from Met1 - Leu71.

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