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

Name: Human/Mouse FoxP3 Antibody
Brand: R&D Systems
SKU: MAB8214
Rating: 3 (1 reviews)

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.

Human/Mouse FoxP3 Antibody

Catalog #: MAB8214
8 Citations 1 Review Datasheet / COA / SDS

Recombinant Monoclonal Antibody

Catalog #	Availability	Size / Price	Qty
MAB8214
MAB8214-SP

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

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Citations (8)

FAQs

Supplemental Products

Reviews (1)

Summary Data Examples Preparation and Storage Reconstitution Calculator Background Product Datasheets Related Research Areas

Human/Mouse FoxP3 Antibody Summary

Species Reactivity

Human, Mouse

Specificity

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

Source

Recombinant Monoclonal Rabbit IgG Clone # 1054C

Purification

Protein A or G purified from cell culture supernatant

Immunogen

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

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Label

Unconjugated

Applications

Recommended Concentration

Sample

Flow Cytometry

0.25 µg/10⁶ cells

See below

Immunohistochemistry

5-25 µg/mL

See below

Dual RNAscope ISH-IHC

5-25 µg/mL

See below

Immunocytochemistry

8-25 µg/mL

See below

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Flow Cytometry

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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).

Flow Cytometry

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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).

Immunocytochemistry

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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.

Immunohistochemistry

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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.

Immunohistochemistry

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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.

Dual RNAscope ISH-IHC Compatible

FoxP3 antibody in Human Tonsil Using Dual RNAscope<sup>®</sup>ISH and IHC.

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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).

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS.

Shipping

The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C

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.

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

Entrez Gene IDs

50943 (Human); 20371 (Mouse)

Alternate Names

AIID; AIIDMGC141961; DIETER; forkhead box P3; Forkhead Box Protein P3; FoxP3; FOXP3delta7; immune dysregulation, polyendocrinopathy, enteropathy, X-linked; Immunodeficiency, Polyendocrinopathy, Enteropathy, X-Linked; IPEX; JM2; MGC141961; MGC141963; PIDX; PIDXMGC141963; SCURFIN; XPID; XPIDpolyendocrinopathy, enteropathy, X-linked

Product Datasheets

Product Datasheet

COA

SDS

Related Research Areas

Citations for Human/Mouse FoxP3 Antibody

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

8 Citations: Showing 1 - 8
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PRSS2 remodels the tumor microenvironment via repression of Tsp1 to stimulate tumor growth and progression
Authors: L Sui, S Wang, D Ganguly, TP El Rayes, C Askeland, A Børretzen, D Sim, OJ Halvorsen, G Knutsvik, J Arnes, S Aziz, S Haukaas, WD Foulkes, DR Bielenberg, A Ziemys, V Mittal, RA Brekken, LA Akslen, RS Watnick
Nature Communications, 2022-12-27;13(1):7959.
Species: Mouse
Sample Types: Whole Tissue
Applications: IHC
Tumor-infiltrated activated B cells suppress liver metastasis of colorectal cancers
Authors: Y Xu, Z Wei, M Feng, D Zhu, S Mei, Z Wu, Q Feng, W Chang, M Ji, C Liu, Y Zhu, L Shen, F Yang, Y Chen, Y Feng, J Xu, D Zhu
Cell Reports, 2022-08-30;40(9):111295.
Species: Mouse
Sample Types: Whole Tissue
Applications: IHC
VEGFR2 activity on myeloid cells mediates immune suppression in the tumor microenvironment
Authors: Y Zhang, H Huang, M Coleman, A Ziemys, P Gopal, SM Kazmi, RA Brekken
JCI Insight, 2021-12-08;6(23):.
Species: Mouse
Sample Types: Whole Tissue
Applications: IHC
Dissecting spatial heterogeneity and the immune-evasion mechanism of CTCs by single-cell RNA-seq in hepatocellular carcinoma
Authors: YF Sun, L Wu, SP Liu, MM Jiang, B Hu, KQ Zhou, W Guo, Y Xu, Y Zhong, XR Zhou, ZF Zhang, G Liu, S Liu, YH Shi, Y Ji, M Du, NN Li, GB Li, ZK Zhao, XY Huang, LQ Xu, QC Yu, DH Peng, SJ Qiu, HC Sun, M Dean, XD Wang, WY Chung, AR Dennison, J Zhou, Y Hou, J Fan, XR Yang
Nature Communications, 2021-07-02;12(1):4091.
Species: Human
Sample Types: Whole Tissue
Applications: IHC
Regulatory T-cell expansion in oral and maxillofacial Langerhans cell histiocytosis
Authors: C Zhang, J Gao, J He, C Liu, X Lv, X Yin, Y Deng, Z Lu, Z Tian
Oral Surg Oral Med Oral Pathol Oral Radiol, 2020-08-08;0(0):.
Species: Human
Sample Types: Whole Tissue
Applications: IHC
Formation of Human Neuroblastoma in Mouse-Human Neural Crest Chimeras
Authors: MA Cohen, S Zhang, S Sengupta, H Ma, GW Bell, B Horton, B Sharma, RE George, S Spranger, R Jaenisch
Cell Stem Cell, 2020-03-05;0(0):.
Species: Mouse
Sample Types: Tissue
Applications: IHC-P
Improved Multiplex Immunohistochemistry for Immune Microenvironment Evaluation of Mouse Formalin-Fixed, Paraffin-Embedded Tissues
Authors: N Sorrelle, D Ganguly, ATA Dominguez, Y Zhang, H Huang, LN Dahal, N Burton, A Ziemys, RA Brekken
J. Immunol., 2018-12-03;0(0):.
Species: Mouse
Sample Types: Whole Tissue
Applications: IHC
CD25 and TGF-? blockade based on predictive integrated immune ratio inhibits tumor growth in pancreatic cancer
Authors: N Pu, G Zhao, H Yin, JA Li, A Nuerxiati, D Wang, X Xu, T Kuang, D Jin, W Lou, W Wu
J Transl Med, 2018-10-25;16(1):294.
Species: Mouse
Sample Types: Whole Tissue
Applications: IHC

FAQs

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

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