Invitrogen

FOXP3 Monoclonal Antibody (FJK-16s), PerCP-eFluor™ 710, eBioscience™

Product Image Gallery

FOXP3 Antibody in Flow Cytometry (Flow) — C57BL/6 mouse splenocytes were stained intracellularly, using the Foxp3/Transcription Factor Staining Buffer Set (Product # 00-5523-00) and protocol, with CD4 Monoclonal Antibody, PE-Cyanine7 (Product # 25-0041-82) and 0.125 µg of Rat IgG2a kappa Isotype Control, PerCP-eFluor 710 (Product # 46-4321-82) (left) or 0.125 µg of Foxp3 Monoclonal Antibody, PerCP-eFluor 710 (right). Cells in the lymphocyte gate were used for analysis.

FOXP3 Antibody in Immunocytochemistry (ICC/IF) — Figure 2 Lower dietary protein intake into middle age results in kidney leukocyte infiltration and inflammation (A) Representative photomicrographs of leukocyte infiltration in periodic acid-Schiff (PAS)-stained kidney cortices according to score 1-3 (x200 magnification) (arrows indicate regions of leukocyte infiltrate). (B) Ordinal regression was used to model the relationship between leukocyte infiltration and macronutrient intake, and plots show the probability of obtaining a score of 1 (red, low infiltration), 2 (green, moderate infiltration), or 3 (blue, high infiltration) as protein (left panel), CHO (middle panel), and fat (right panel) intake increased (n = 160). (C) Kidney leukocyte infiltration scores presented by tertiles of dietary protein intake (low < 6.5 kJ/mouse/day, med 6.5-12 kJ/mouse/day, high 12.1-30 kJ/mouse/day). (D) Kidney leukocyte infiltration scores presented by tertiles of dietary fat intake (low <8.58 kJ/mouse/day, med 8.59-13.8 kJ/mouse/day, high 13.8-44.9 kJ/mouse/day). (E) Representative photomicrographs of regions of the renal cortex with leukocyte infiltrate (x800) stained for T cell markers CD3, CD4, FoxP3. (F) Summarized p values for coefficients for the GAMs of kidney cytokine profiling using Legendplex 13-plex pro-inflammatory cytokine array. p values for individual effects (single column) or interactive effects (spanning 2 or more columns) of macronutrients on cortical cytokine profiles are shown (NS, non-significant; n numbers and full p

FOXP3 Antibody in Immunohistochemistry (IHC) — Figure 4 Characterization of T cell population in the kidneys of 19 and 15-week-old female MRL-Fas lpr KSRP -/- mice compared to MRL-Fas lpr KSRP +/+ mice ( a ) RNA was isolated from the kidneys of 19-week-old female MRL-Fas lpr KSRP -/- mice (KO) and MRL-Fas lpr KSRP +/+ mice (WT). T cell lineage-specific transcription factor mRNA expression of T-bet, GATA3, RORgammaT and FoxP3 were analyzed by qRT-PCR and normalized to the mRNA expression of Pol2a. Data presented are the mean +- SEM of the relative mRNA expressions based on WT female 19 weeks (100%) 9 to 14 animals per genotype (*: p < 0.05; n.s. : not significantly different from WT, T -TEST). ( b ) Immunostaining was used to analyze intra-renal frequency of FoxP3, Ki67 and IFN-gamma producing cells in the kidneys of female 19-week-old MRL-Fas pr KSRP +/+ and MRL-Fas lpr KSRP -/- mice. Data presented are the mean +- SEM of the score with 10 animals per genotype (**: p < 0.01, *: p < 0.05 compared to WT, TTEST). Representative image of immunostaining with 40x magnification. ( c - e ) FACS was used to analyze intra-renal frequency of CD4 + , CD4 + -IFN-gamma + and FoxP3 + cells in the kidneys of female 15-week-old MRL-Fas lpr KSRP +/+ and MRL-Fas pr KSRP -/- mice. Data presented are the mean +- SEM of the relative cell numbers compared to MRL-Fas lpr KSRP +/+ mice (19 to 23 animals per genotype) (**: p < 0.01, n.s. : not significant, compared to WT, t -test).

Product Details

46-5773-82

Applications	Tested Dilution	Publications
Flow Cytometry (Flow)	0.25 µg/test	View 52 publications 52 publications
Immunocytochemistry (ICC/IF)	-	View 1 publication 1 publication
Immunohistochemistry (IHC)	-	View 10 publications 10 publications

Product Specifications
Species Reactivity	Bovine, Dog, Cat, Mouse, Pig, Rat
Published species	Not Applicable
Host/Isotype	Rat / IgG2a, kappa
Recommended Isotype Control	Rat IgG2a kappa Isotype Control (eBR2a), PerCP-eFluor™ 710, eBioscience™
Class	Monoclonal
Type	Antibody
Clone	FJK-16s
Conjugate	PerCP-eFluor™ 710 Alexa Fluor 488 Alexa Fluor 532 Alexa Fluor 561 Alexa Fluor 660 Alexa Fluor 700 APC Biotin Brilliant Violet 421 eFluor 450 eFluor 506 eFluor 570 eFluor 615 eFluor 660 FITC PE PE-Cyanine5 PE-Cyanine5.5 PE-Cyanine7 PE-eFluor 610 PerCP-Cyanine5.5 Unconjugated
Excitation/Emission Max	482/708 nm View spectra
Form	Liquid
Concentration	0.2 mg/mL
Purification	Affinity chromatography
Storage buffer	PBS, pH 7.2
Contains	0.09% sodium azide
Storage conditions	4° C, store in dark, DO NOT FREEZE!
RRID	AB_2811810

Product Specific Information

Description: The FJK-16s antibody reacts with mouse, rat, dog, porcine, bovine and cat Foxp3 also known as FORKHEAD BOX P3, SCURFIN, and JM2; cross reactivity of this antibody to other proteins has not been determined. Foxp3, a 49-55 kDa protein, is a member of the forkhead/winged-helix family of transcriptional regulators, and was identified as the gene defective in 'scurfy' (sf) mice. Constitutive high expression of foxP3 mRNA has been shown in CD4+CD25+ regulatory T cells (Treg cells), and ectopic expression of foxp3 in CD4+CD25- cells imparts a Treg phenotype in these cells.

Immunoblotting with FJK-16s antibody has mapped the epitope to amino acids 75-125 of the mouse Foxp3 protein. In the human, this region has been shown to be alternatively spliced at the mRNA level. Both the alternatively-spliced and non-spliced isoforms are present in the CD4+CD25+ subset of lymphocytes. Preliminary RT-PCR experiments have not revealed this alternatively-spliced isoform in mouse splenocytes, suggesting different gene regulation in the mouse and human.

Please note that FJK-16s has been optimized for use with the Foxp3/Transcription Factor Buffer Staining Set (cat. 00-5523). The use of other fixation and staining buffers is not recommended.

Applications Reported: This FJK-16s antibody has been reported for use in intracellular staining followed by flow cytometric analysis.

Applications Tested: This FJK-16s antibody has been tested by intracellular staining and flow cytometric analysis of mouse splenocytes using the Foxp3/Transcription Factor Staining Buffer Set (Product # 00-5523-00) and protocol. Please refer to "Staining Intracellular Antigens for Flow Cytometry, Protocol B: One step protocol for intracellular (nuclear) proteins" located at www.thermofisher.com/flowprotocols. This may be used at less than or equal to 0.25 µg per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.

PerCP-eFluor 710 emits at 710 nm and is excited with the blue laser (488 nm); it can be used in place of PerCP-Cyanine5.5. We recommend using a 710/50 bandpass filter, however, the 695/40 bandpass filter is an acceptable alternative. Please make sure that your instrument is capable of detecting this fluorochrome.

Light sensitivity: This tandem dye is sensitive to photo-induced oxidation. Please protect this vial and stained samples from light.

Fixation: Samples can be stored in IC Fixation Buffer (Product # 00-8222-49) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 00-5333-57) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically.

Excitation: 488 nm; Emission: 710 nm; Laser: Blue Laser

Target Information

FOXP3 (Forkhead box protein 3) is a member of the forkhead/winged-helix family of transcriptional regulators, highly conserved across mammals, and essential for normal immune homeostasis. FOXP3 is 381 amino acids long, stably and constitutively expressed at a high level in CD25 + CD4 positive regulatory T cells, a low level in CD4-positive/CD25-negative cells, and is absent in CD4-negative/CD8-positive T cells. FOXP3 may be a master regulatory gene, and a more specific marker of regulatory T cells. Defects in the gene encoding FOXP3 protein cause the scurfy phenotype in mice. In humans FOXP3 defects play a role in IPEX syndrome (immune dysfunction, polyendocrinopathy, enteropathy, X-linked syndrome), also known as X-linked autoimmunity-allergic dysregulation (XLAAD) syndrome. Transcript variants of FOXP3 encoding different isoforms have been identified. In human breast and colon cancer cells, expression of FOXP3 is regulated by p53 in response to the DNA damage.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

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JCI insight

Zammit NW,Walters SN,Seeberger KL,O'Connell PJ,Korbutt GS,Grey ST

Published figure using FOXP3 monoclonal antibody (Product # 46-5773-82) in Flow Cytometry

Fri Nov 01 00:00:00 EDT 2019

Targeted knock-in mice expressing the oxidase-fixed form of xanthine oxidoreductase favor tumor growth.

Nature communications

Kusano T,Ehirchiou D,Matsumura T,Chobaz V,Nasi S,Castelblanco M,So A,Lavanchy C,Acha-Orbea H,Nishino T,Okamoto K,Busso N

Published figure using FOXP3 monoclonal antibody (Product # 46-5773-82) in Flow Cytometry

Mon Oct 28 00:00:00 EDT 2019

Antitumor and immunomodulatory effects of a novel multitarget inhibitor, CS2164, in mouse hepatocellular carcinoma models.

Anti-cancer drugs

Zhou Y,Fu C,Kong Y,Pan D,Wang Y,Huang S,Li Z,Ning Z,Lu X,Shan S,Xin L

Published figure using FOXP3 monoclonal antibody (Product # 46-5773-82) in Flow Cytometry

Tue Oct 01 00:00:00 EDT 2019

Hijacking antibody-induced CTLA-4 lysosomal degradation for safer and more effective cancer immunotherapy.

Cell research

Zhang Y,Du X,Liu M,Tang F,Zhang P,Ai C,Fields JK,Sundberg EJ,Latinovic OS,Devenport M,Zheng P,Liu Y

Published figure using FOXP3 monoclonal antibody (Product # 46-5773-82) in Flow Cytometry

Thu Aug 01 00:00:00 EDT 2019

Loss of TET2 and TET3 in regulatory T cells unleashes effector function.

Nature communications

Yue X,Lio CJ,Samaniego-Castruita D,Li X,Rao A

Published figure using FOXP3 monoclonal antibody (Product # 46-5773-82) in Flow Cytometry

Wed May 01 00:00:00 EDT 2019

Anti-IL-6 eluting immunomodulatory biomaterials prolong skin allograft survival.

Scientific reports

Uehara M,Li X,Sheikhi A,Zandi N,Walker B,Saleh B,Banouni N,Jiang L,Ordikhani F,Dai L,Yonar M,Vohra I,Kasinath V,Orgill DP,Khademhosseini A,Annabi N,Abdi R

Published figure using FOXP3 monoclonal antibody (Product # 46-5773-82) in Flow Cytometry

Thu Apr 25 00:00:00 EDT 2019

MTOR signaling is essential for the development of thymic epithelial cells and the induction of central immune tolerance.

Autophagy

Liang Z,Zhang L,Su H,Luan R,Na N,Sun L,Zhao Y,Zhang X,Zhang Q,Li J,Zhang L,Zhao Y

Published figure using FOXP3 monoclonal antibody (Product # 46-5773-82) in Flow Cytometry

Fri Mar 22 00:00:00 EDT 2019

Characterization of immune cell subtypes in three commonly used mouse strains reveals gender and strain-specific variations.

Laboratory investigation; a journal of technical methods and pathology

Hensel JA,Khattar V,Ashton R,Ponnazhagan S

Published figure using FOXP3 monoclonal antibody (Product # 46-5773-82) in Flow Cytometry

Tue Jan 01 00:00:00 EST 2019

REV-ERBα Regulates T_H17 Cell Development and Autoimmunity.

Cell reports

Amir M,Chaudhari S,Wang R,Campbell S,Mosure SA,Chopp LB,Lu Q,Shang J,Pelletier OB,He Y,Doebelin C,Cameron MD,Kojetin DJ,Kamenecka TM,Solt LA

Published figure using FOXP3 monoclonal antibody (Product # 46-5773-82) in Flow Cytometry

Wed Dec 26 00:00:00 EST 2018

Control of T_reg cell homeostasis and immune equilibrium by Lkb1 in dendritic cells.

Nature communications

Chen S,Fang L,Guo W,Zhou Y,Yu G,Li W,Dong K,Liu J,Luo Y,Wang B,Li Z,Zhao C,Sun Z,Shen Y,Leng Q,Zhou D,Han Z,Huang H,Ren H,Xu G,Feng X

Published figure using FOXP3 monoclonal antibody (Product # 46-5773-82) in Flow Cytometry

Thu Dec 13 00:00:00 EST 2018

Hippo Kinases Mst1 and Mst2 Sense and Amplify IL-2R-STAT5 Signaling in Regulatory T Cells to Establish Stable Regulatory Activity.

Immunity

Shi H,Liu C,Tan H,Li Y,Nguyen TM,Dhungana Y,Guy C,Vogel P,Neale G,Rankin S,Feng Y,Peng J,Tao W,Chi H

Published figure using FOXP3 monoclonal antibody (Product # 46-5773-82) in Flow Cytometry

Tue Nov 20 00:00:00 EST 2018

Indirect presentation in the thymus limits naive and regulatory T-cell differentiation by promoting deletion of self-reactive thymocytes.

Immunology

Yap JY,Wirasinha RC,Chan A,Howard DR,Goodnow CC,Daley SR

Published figure using FOXP3 monoclonal antibody (Product # 46-5773-82) in Flow Cytometry

Sun Jul 01 00:00:00 EDT 2018

Mitophagy in Intestinal Epithelial Cells Triggers Adaptive Immunity during Tumorigenesis.

Cell

Ziegler PK,Bollrath J,Pallangyo CK,Matsutani T,Canli Ö,De Oliveira T,Diamanti MA,Müller N,Gamrekelashvili J,Putoczki T,Horst D,Mankan AK,Öner MG,Müller S,Müller-Höcker J,Kirchner T,Slotta-Huspenina J,Taketo MM,Reinheckel T,Dröse S,Larner AC,Wels WS,Ernst M,Greten TF,Arkan MC,Korn T,Wirth D,Greten FR

Published figure using FOXP3 monoclonal antibody (Product # 46-5773-82) in Flow Cytometry

Thu Jun 28 00:00:00 EDT 2018

The effects of freeze-dried Ganoderma lucidum mycelia on a recurrent oral ulceration rat model.

BMC complementary and alternative medicine

Xie L,Zhong X,Liu D,Liu L,Xia Z

Published figure using FOXP3 monoclonal antibody (Product # 46-5773-82) in Flow Cytometry

Fri Dec 01 00:00:00 EST 2017

A bidirectional association between the gut microbiota and CNS disease in a biphasic murine model of multiple sclerosis.

Gut microbes

Colpitts SL,Kasper EJ,Keever A,Liljenberg C,Kirby T,Magori K,Kasper LH,Ochoa-Repáraz J

Published figure using FOXP3 monoclonal antibody (Product # 46-5773-82) in Flow Cytometry

Thu Nov 02 00:00:00 EDT 2017

Bioinformatics

Protein Aliases: Forkhead box protein P3; Forkhead box protein P3 41 kDa form; Forkhead box protein P3, C-terminally processed; forkhead/winged helix transcription factor 3; Foxp3; MGC141961; MGC141963; RP23-54C14.1; Scurfin; scurfy

Gene Aliases: Foxp3; JM2; RGD1562112; scurfin; sf

UniProt ID: (Mouse) Q99JB6

Entrez Gene ID: (Dog) 491876, (Pig) 444998, (Bovine) 506053, (Cat) 100037405, (Mouse) 20371, (Rat) 317382

Function(s)

Process(es)

core promoter binding nucleic acid binding DNA binding transcription factor activity, sequence-specific DNA binding transcription factor activity, RNA polymerase II distal enhancer sequence-specific binding transcription corepressor activity protein binding histone acetyltransferase binding protein homodimerization activity histone deacetylase binding sequence-specific DNA binding metal ion binding NFAT protein binding RNA polymerase II transcription factor activity, sequence-specific DNA binding molecular_function

FOXP3 Monoclonal Antibody (FJK-16s), PerCP-eFluor™ 710, eBioscience™

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