Invitrogen

FOXP3 Monoclonal Antibody (150D/E4), eBioscience™

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FOXP3 Antibody in Western Blot (WB) — Published figure using FOXP3 monoclonal antibody (Product # 14-4774-82) in Western Blot

FOXP3 Antibody in Western Blot, Flow Cytometry (WB, Flow) — Published figure using FOXP3 monoclonal antibody (Product # 14-4774-82) in Western Blot and Flow Cytometry

FOXP3 Antibody in Flow Cytometry (Flow) — Fig. 6 TRA expression by distinct mTEC compartments impacts the mode of tolerance induction in the polyclonal T cell repertoire in LCMV infected mice. Mice were infected with LCMV-OVA and the antigen-specific response was measured in the spleen 8 days post-infection (NI, not infected mice) a, c Representative CD44 and H-2K b -OVA or H-2D b -gp33 tetramer staining on CD8 + T cells from littermate controls (LMC), CRP, and INS2 Tg mice. Relative response and absolute number of OVA- ( b ) and gp33- ( d ) specific CD8 + T cell response in USA expressing mice as compared to LMC. e Representative CD44 and I-A b -gp66 tetramer staining on CD4 + T cells from LMC, CRP, and INS2 Tg mice. f Relative and absolute gp66-specific response in USA expressing mice. g Representative flow plot of CD25 and FoxP3, gated on live I-A b -gp66-specific CD4 + T cells. h Relative proportion of CD25 + FoxP3 + T reg among gp66-specific CD4 + T cells and absolute number of gp66-specific CD4 + CD25 + FoxP3 + T reg (mean +- SEM, b left panel n = 22 LMC, 7 CRP, 5 CRP lo , 9 INS2, 3 INS2 lo ; b right panel n = 3 NI, 8 CRP LMC, 10 CRP Tg, 9 CRP lo LMC, 9 CRP lo Tg, 10 INS2 LMC, 9 INS2 Tg, 3 INS2 lo LMC, 3 INS2 lo Tg; d and f left panel n = 32 LMC, 12 CRP, 12 CRP lo , 14 INS2, 8 INS2 lo , d right panel n = 6 NI, 11 CRP LMC, 12 CRP Tg, 11 CRP lo LMC, 12 CRP lo Tg, 13 INS2 LMC, 11 INS2 Tg, 6 INS2 lo LMC, 6 INS2 lo Tg, f right panel n = 6 NI, 11 CRP LMC, 13 CRP Tg, 12 CRP lo LMC, 13 CRP lo Tg, 14 INS2 LMC, 12 INS2 T

FOXP3 Antibody in ChIP assay (ChIP) — Figure 3 Conditional HDAC11 deletion promotes chromatin remodeling at the Foxp3 locus and Treg gene expression. ( A ) ChIP studies of the Foxp3 promoter and 3 enhancer regions in WT vs. conditional HDAC11-/- Treg cells, showing increased acetyl-H3 binding at all 4 sites in HDAC11-/- vs. WT Treg cells. ( B ) Luciferase reporter assays showing the impact of overexpression, and also inhibition, of HDAC11 in 293 T cells that were transiently transfected with murine Foxp3-promoter-Luc vector plus expression vectors for p65, HDAC11, Foxp3, and p300, as specified, or empty vector (EV) along with 50 ng of pRL-TK Renilla luciferase internal control vector; EV was added to ensure equal amounts of DNA per reaction. After 48 h, cells were treated with PMA (6 ng/ml) and ionomycin (1 muM) for 5 to 6 h, and 1 muM HDAC11i where specified, and cell lysates were harvested for luciferase assays. Firefly luminescence data was normalized to Renilla luminescence and analyzed by ANOVA (*p < 0.05, **p < 0.005); data are representative of the results of 3 or more independent experiments. ( C ) Heat-map of selected genes that were upregulated (red) or downregulated (blue) in conditional HDAC11-/- vs. WT Treg cells. ( D ) qPCR evaluation of WT vs. conditional HDAC11-/- Treg cells, showing increased expression of Jun, Junb, Fos and Fosb in HDAC11-/- Tregs; normalized data are shown as mean +- SD, 4 mice/group, and *p < 0.05, **p < 0.01.

Product Details

14-4774-82

Applications	Tested Dilution	Publications
Western Blot (WB)	Assay-Dependent	View 9 publications 9 publications
Immunohistochemistry (IHC)	Assay-Dependent	-
Immunohistochemistry (Paraffin) (IHC (P))	-	View 1 publication 1 publication
Flow Cytometry (Flow)	-	View 10 publications 10 publications
Immunoprecipitation (IP)	Assay-Dependent	View 1 publication 1 publication
ChIP assay (ChIP)	-	View 2 publications 2 publications

Product Specifications
Species Reactivity	Human, Mouse, Rat
Published species	Human, Mouse
Host/Isotype	Mouse / IgG1, kappa
Class	Monoclonal
Type	Antibody
Clone	150D/E4
Conjugate	Unconjugated Alexa Fluor 488 eFluor 660 PE
Form	Liquid
Concentration	0.5 mg/mL
Purification	Affinity chromatography
Storage buffer	PBS, pH 7.2
Contains	0.09% sodium azide
Storage conditions	4° C
RRID	AB_467552

Product Specific Information

Description: eBioscience offers a panel of monoclonal antibodies to different epitopes of human/primate and mouse/rat Foxp3 protein, providing useful tools for investigating the complete expression pattern of Foxp3 at the protein level, and discerning the precise subsets of Foxp3+ cells. Other antibodies to Foxp3 available from eBioscience, which have been used significantly in scientific literature, include the anti-human Foxp3 PCH101 (cat. 72-5776) and ebio7979 (cat. 12-7979), and the anti-mouse/rat Foxp3 FJK-16s (cat. 72-5775). The 150D/E4 has been mapped to the splice region in Exon 2 found in human cells. The splicing of this region has not been shown to occur in mouse. Characterization of this antibody to the splice variant has been limited to epitope mapping. It should be noted that other antibodies such as PCH101 and 236A/E7 will recognize both the spliced and full length forms of the Foxp3 protein and stain more intensely than 150D/E4 and FJK-16s in human cells.

The 150D/E4 antibody reacts with mouse/rat/human 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.

Applications Reported: This 150D/E4 antibody has been reported for use in immunoblotting (WB), and immunohistochemical staining.

Applications Tested: This 150D/E4 antibody has been tested by immunoblotting of recombinant mouse and human Foxp3 protein.

The purified format is not recommended for intracellular staining and flow cytometric analysis.

Purity: Greater than 90%, as determined by SDS-PAGE.

Aggregation: Less than 10%, as determined by HPLC.

Filtration: 0.2 µm post-manufacturing filtered.

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.

Product References

AMP-activated protein kinase alpha1 promotes tumor development via FOXP3 elevation in tumor-infiltrating Treg cells.

iScience

An J,Ding Y,Yu C,Li J,You S,Liu Z,Song P,Zou MH

14-4774-82 was used in Western Blot to suggest that AMPKα1 activation promotes tumor growth by maintaining FOXP3 stability in tumor-infiltrating Treg cells and that selective inhibition of AMPK in Treg cells might be an effective anti-tumor therapy.

Fri Jan 21 00:00:00 EST 2022

MEF2D sustains activation of effector Foxp3+ Tregs during transplant survival and anticancer immunity.

The Journal of clinical investigation

Di Giorgio E,Wang L,Xiong Y,Akimova T,Christensen LM,Han R,Samanta A,Trevisanut M,Bhatti TR,Beier UH,Hancock WW

14-4774 was used in Western Blotting to study the role played by MEF2D in sustaining effector Foxp3+ Treg functions without abrogating their basal actions suggests its suitability for drug discovery efforts in cancer therapy.

Tue Dec 01 00:00:00 EST 2020

Differential expression of tissue-restricted antigens among mTEC is associated with distinct autoreactive T cell fates.

Nature communications

Lebel MÈ,Coutelier M,Galipeau M,Kleinman CL,Moon JJ,Melichar HJ

Published figure using FOXP3 monoclonal antibody (Product # 14-4774-82) in Flow Cytometry

Fri Jul 24 00:00:00 EDT 2020

Inhibiting the coregulator CoREST impairs Foxp3+ Treg function and promotes antitumor immunity.

The Journal of clinical investigation

Xiong Y,Wang L,Di Giorgio E,Akimova T,Beier UH,Han R,Trevisanut M,Kalin JH,Cole PA,Hancock WW

14-4774 was used in Western Blotting to analyse the role in Tregs of the evolutionarily conserved CoREST complex, consisting of a scaffolding protein, Rcor1 or Rcor2, plus Hdac1 or Hdac2 and Lsd1 enzymes.

Wed Apr 01 00:00:00 EDT 2020

Mitochondrial transfer from MSCs to T cells induces Treg differentiation and restricts inflammatory response.

EMBO reports

Court AC,Le-Gatt A,Luz-Crawford P,Parra E,Aliaga-Tobar V,Bátiz LF,Contreras RA,Ortúzar MI,Kurte M,Elizondo-Vega R,Maracaja-Coutinho V,Pino-Lagos K,Figueroa FE,Khoury M

Published figure using FOXP3 monoclonal antibody (Product # 14-4774-82) in Flow Cytometry

Wed Feb 05 00:00:00 EST 2020

Alternative Splicing of FOXP3 Controls Regulatory T Cell Effector Functions and Is Associated With Human Atherosclerotic Plaque Stability.

Circulation research

Joly AL,Seitz C,Liu S,Kuznetsov NV,Gertow K,Westerberg LS,Paulsson-Berne G,Hansson GK,Andersson J

14-4774 was used in Flow cytometry/Cell sorting to define the regulation and functions of the 2 major FOXP3 isoforms, FOXP3fl and FOXP3Δ2, as well as to establish whether their expression is associated with the ischemic atherosclerotic disease.

Fri May 11 00:00:00 EDT 2018

Histone/protein deacetylase 11 targeting promotes Foxp3+ Treg function.

Scientific reports

Huang J,Wang L,Dahiya S,Beier UH,Han R,Samanta A,Bergman J,Sotomayor EM,Seto E,Kozikowski AP,Hancock WW

Published figure using FOXP3 monoclonal antibody (Product # 14-4774-82) in ChIP assay

Thu Aug 17 00:00:00 EDT 2017

Histone/protein deacetylase 11 targeting promotes Foxp3+ Treg function.

Scientific reports

Huang J,Wang L,Dahiya S,Beier UH,Han R,Samanta A,Bergman J,Sotomayor EM,Seto E,Kozikowski AP,Hancock WW

Published figure using FOXP3 monoclonal antibody (Product # 14-4774-82) in ChIP assay

Thu Aug 17 00:00:00 EDT 2017

Foxp3 Reprograms T Cell Metabolism to Function in Low-Glucose, High-Lactate Environments.

Cell metabolism

Angelin A,Gil-de-Gómez L,Dahiya S,Jiao J,Guo L,Levine MH,Wang Z,Quinn WJ,Kopinski PK,Wang L,Akimova T,Liu Y,Bhatti TR,Han R,Laskin BL,Baur JA,Blair IA,Wallace DC,Hancock WW,Beier UH

14-4774 was used in Chromatin immunoprecipitation to report that the Treg transcription factor Foxp3 reprograms T cell metabolism by suppressing Myc and glycolysis.

Tue Jun 06 00:00:00 EDT 2017

Cutting Edge: Increased Autoimmunity Risk in Glycogen Storage Disease Type 1b Is Associated with a Reduced Engagement of Glycolysis in T Cells and an Impaired Regulatory T Cell Function.

Journal of immunology (Baltimore, Md. : 1950)

Melis D,Carbone F,Minopoli G,La Rocca C,Perna F,De Rosa V,Galgani M,Andria G,Parenti G,Matarese G

14477482 was used in western blot to examine the effect of glucose-6-phosphate transporter mutation on immune cell homeostasis and CD4+ T cell functions

Mon May 15 00:00:00 EDT 2017

Microbial, metabolomic, and immunologic dynamics in a relapsing genetic mouse model of colitis induced by T-synthase deficiency.

Gut microbes

Jacobs JP,Lin L,Goudarzi M,Ruegger P,McGovern DP,Fornace AJ,Borneman J,Xia L,Braun J

Published figure using FOXP3 monoclonal antibody (Product # 14-4774-82) in Flow Cytometry

Mon Jan 02 00:00:00 EST 2017

Differential Role of Hematopoietic and Nonhematopoietic Cell Types in the Regulation of NK Cell Tolerance and Responsiveness.

Journal of immunology (Baltimore, Md. : 1950)

Shifrin NT,Kissiov DU,Ardolino M,Joncker NT,Raulet DH

14477482 was used in flow cytometry to study the role of MCH I in natural killer cell tolerance and hyporesponsiveness

Tue Nov 15 00:00:00 EST 2016

Characteristics of CD4⁺CD25⁺Foxp3⁺ regulatory T cells in patients with multiple organ dysfunction syndrome.

Experimental and therapeutic medicine

Zang X,Jiang Y,Li X,DU Y,Niu J

Published figure using FOXP3 monoclonal antibody (Product # 14-4774-82) in Flow Cytometry

Sun May 01 00:00:00 EDT 2016

TGF-Beta Blockade Increases Renal Inflammation Caused by the C-Terminal Module of the CCN2.

Mediators of inflammation

Rodrigues-Díez R,Rayego-Mateos S,Orejudo M,Aroeira LS,Selgas R,Ortiz A,Egido J,Ruiz-Ortega M

Fri Feb 12 00:00:00 EST 2016

Glycolysis controls the induction of human regulatory T cells by modulating the expression of FOXP3 exon 2 splicing variants.

Nature immunology

De Rosa V,Galgani M,Porcellini A,Colamatteo A,Santopaolo M,Zuchegna C,Romano A,De Simone S,Procaccini C,La Rocca C,Carrieri PB,Maniscalco GT,Salvetti M,Buscarinu MC,Franzese A,Mozzillo E,La Cava A,Matarese G

Published figure using FOXP3 monoclonal antibody (Product # 14-4774-82) in Flow Cytometry

Sun Nov 01 00:00:00 EDT 2015

Glycolysis controls the induction of human regulatory T cells by modulating the expression of FOXP3 exon 2 splicing variants.

Nature immunology

Published figure using FOXP3 monoclonal antibody (Product # 14-4774-82) in Flow Cytometry

Sun Nov 01 00:00:00 EDT 2015

Glycolysis controls the induction of human regulatory T cells by modulating the expression of FOXP3 exon 2 splicing variants.

Nature immunology

Published figure using FOXP3 monoclonal antibody (Product # 14-4774-82) in Flow Cytometry

Sun Nov 01 00:00:00 EDT 2015

The C-terminal module IV of connective tissue growth factor is a novel immune modulator of the Th17 response.

Laboratory investigation; a journal of technical methods and pathology

Rodrigues-Díez R,Rodrigues-Díez RR,Rayego-Mateos S,Suarez-Alvarez B,Lavoz C,Stark Aroeira L,Sánchez-López E,Orejudo M,Alique M,Lopez-Larrea C,Ortiz A,Egido J,Ruiz-Ortega M

14-4774 was used in Western Blotting to reveal that CCN2's C-terminal module induces Th17 differentiation of human Th17 cells and causes a renal Th17 inflammatory response.

Mon Jul 01 00:00:00 EDT 2013

CD40 deficiency in mice exacerbates obesity-induced adipose tissue inflammation, hepatic steatosis, and insulin resistance.

American journal of physiology. Endocrinology and metabolism

Guo CA,Kogan S,Amano SU,Wang M,Dagdeviren S,Friedline RH,Aouadi M,Kim JK,Czech MP

Published figure using FOXP3 monoclonal antibody (Product # 14-4774-82) in Flow Cytometry

Wed May 01 00:00:00 EDT 2013

Paradoxically increased FOXP3+ T cells in IBD do not preferentially express the isoform of FOXP3 lacking exon 2.

Digestive diseases and sciences

Lord JD,Valliant-Saunders K,Hahn H,Thirlby RC,Ziegler SF

14-4774 was used in Immunohistochemistry on paraffin embedded tissues to investigate whether IBD patients preferentially express the δexon2 variant of FOXP3.

Thu Nov 01 00:00:00 EDT 2012

Protein kinase B/Akt signals impair Th17 differentiation and support natural regulatory T cell function and induced regulatory T cell formation.

Journal of immunology (Baltimore, Md. : 1950)

Pierau M,Engelmann S,Reinhold D,Lapp T,Schraven B,Bommhardt UH

14-4774 was used in Western Blotting to indicate an important role of PKB signals on control of TGF-beta1-mediated T cell responses and, thereby, on tolerizing and inflammatory immune processes.

Sun Nov 15 00:00:00 EST 2009

Critical roles of lysosomal acid lipase in T cell development and function.

The American journal of pathology

Qu P,Du H,Wilkes DS,Yan C

14-4774-82 was used in Flow Cytometry to conclude that the altered development and function of lal(-/-) T cells contributes to disease formation in various organs during LAL deficiency.

Sun Mar 01 00:00:00 EST 2009

Malignant Tregs express low molecular splice forms of FOXP3 in Sézary syndrome.

Leukemia

Krejsgaard T,Gjerdrum LM,Ralfkiaer E,Lauenborg B,Eriksen KW,Mathiesen AM,Bovin LF,Gniadecki R,Geisler C,Ryder LP,Zhang Q,Wasik MA,Odum N,Woetmann A

14-4774 was used in Western Blotting to investigate the underlying mechanism of immunodeficiency during Sézary syndrome.

Mon Dec 01 00:00:00 EST 2008

Epigenetic inheritance of DNA methylation limits activation-induced expression of FOXP3 in conventional human CD25-CD4+ T cells.

International immunology

Nagar M,Vernitsky H,Cohen Y,Dominissini D,Berkun Y,Rechavi G,Amariglio N,Goldstein I

14-4774 was used in Flow cytometry/Cell sorting to investigate the mode of de novo activation-dependent expression of FOXP3 in conventional human CD4+ T cells.

Fri Aug 01 00:00:00 EDT 2008

Bioinformatics

Protein Aliases: Forkhead box protein P3; Forkhead box protein P3 41 kDa form; Forkhead box protein P3, C-terminally processed; FOXP3delta7; immune dysregulation, polyendocrinopathy, enteropathy, X-linked; immunodeficiency, polyendocrinopathy, enteropathy, X-linked; MGC141961; MGC141963; RP23-54C14.1; Scurfin; scurfy

Gene Aliases: AIID; DIETER; FOXP3; IPEX; JM2; PIDX; RGD1562112; scurfin; sf; XPID

UniProt ID: (Human) Q9BZS1, (Mouse) Q99JB6

Entrez Gene ID: (Human) 50943, (Mouse) 20371, (Rat) 317382

Function(s)