Invitrogen

FOXP3 Monoclonal Antibody (PCH101), Alexa Fluor™ 660, eBioscience™

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Protocols
Questions & Answers
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Product Details

606-4776-42

Applications
Tested Dilution
Publications

Flow Cytometry (Flow)

5 µL (0.125 µg)/test
-
Product Specifications

Species Reactivity

Chimpanzee, Cynomolgus monkey, Human, Non-human primate, Rhesus monkey

Host/Isotype

Rat / IgG2a, kappa

Recommended Isotype Control

Rat IgG2a kappa Isotype Control (eBR2a), Alexa Fluor™ 660, eBioscience™

Class

Monoclonal

Type

Antibody

Clone

PCH101

Conjugate

Alexa Fluor™ 660 Alexa Fluor™ 660 Alexa Fluor™ 660

Excitation/Emission Max

663/691 nm View spectra spectra

Form

Liquid

Concentration

5 µL/Test

Purification

Affinity chromatography

Storage buffer

PBS, pH 7.2, with 0.2% BSA

Contains

0.09% sodium azide

Storage conditions

4° C, store in dark, DO NOT FREEZE!

Shipping conditions

Ambient (domestic); Wet ice (international)

RRID

AB_2896277

Product Specific Information

Description: eBioscience offers a panel of monoclonal antibodies to different epitopes of human Foxp3, providing useful tools for investigating the complete expression pattern of Foxp3 at the protein level, and discerning the precise subsets of Foxp3+ cells.

The PCH101 antibody reacts with the amino terminus of human foxp3 protein 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.

Intracellular staining of human peripheral blood mononuclear cells (PBMCs) with PCH101 antibody using the anti-human Foxp3 Staining Set and protocol reveals approximately 0.5-4% of lymphocytes staining, with the majority of staining occurring in the CD25-bright population. This is subject to donor variability.

PCH101 crossreacts with rhesus, chimpanzee and cynomolgus. We recommend the use of CD4 (OKT4, Product # 11-0048-42 , or RPA-T4, Product # 11-0049-42 , depending on the species) and CD25 (BC96, Product # 17-0259-42).

Applications Reported: This PCH101 antibody has been reported for use in intracellular staining followed by flow cytometric analysis.

Applications Tested: This PCH101 antibody has been pre-diluted and tested by intracellular staining followed by flow cytometric analysis of normal human peripheral blood cells 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 Flow Protocols . This may be used at 5 µL (0.125 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.

Alexa Fluor 660 emits at 690 nm and is intended for use on spectral cytometers where it may be multiplexed with APC, Alexa Fluor 647 and Alexa Fluor 700.

Excitation: 662 nm; Emission: 690 nm; Laser: Red 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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Bioinformatics

Protein Aliases: Forkhead box protein P3; FOXP3delta7; immune dysregulation, polyendocrinopathy, enteropathy, X-linked; immunodeficiency, polyendocrinopathy, enteropathy, X-linked; MGC141961; MGC141963; regulatory protein Foxp3; RP23-54C14.1; Scurfin

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Gene Aliases: AIID; DIETER; FOXP3; IPEX; JM2; PIDX; XPID

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UniProt ID: (Human) Q9BZS1, (Cynomolgus monkey) Q6U8D7

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Entrez Gene ID: (Human) 50943, (Cynomolgus monkey) 102120882, (Rhesus monkey) 574303

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Function(s)
RNA polymerase II transcription factor activity, sequence-specific DNA binding core promoter 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 NF-kappaB binding NFAT protein binding
Process(es)
negative regulation of transcription from RNA polymerase II promoter B cell homeostasis cytokine production myeloid cell homeostasis CD4-positive, CD25-positive, alpha-beta regulatory T cell lineage commitment T cell mediated immunity tolerance induction to self antigen regulation of T cell anergy positive regulation of T cell anergy negative regulation of chronic inflammatory response negative regulation of T cell cytokine production positive regulation of peripheral T cell tolerance induction chromatin remodeling transcription, DNA-templated regulation of transcription, DNA-templated negative regulation of cell proliferation response to virus negative regulation of histone deacetylation negative regulation of NF-kappaB transcription factor activity negative regulation of interferon-gamma production negative regulation of interleukin-10 production negative regulation of interleukin-17 production negative regulation of interleukin-2 production negative regulation of interleukin-4 production negative regulation of interleukin-5 production negative regulation of interleukin-6 production negative regulation of tumor necrosis factor production positive regulation of interleukin-4 production negative regulation of CREB transcription factor activity positive regulation of CD4-positive, CD25-positive, alpha-beta regulatory T cell differentiation positive regulation of transforming growth factor beta1 production positive regulation of immature T cell proliferation in thymus positive regulation of histone acetylation negative regulation of histone acetylation negative regulation of cytokine biosynthetic process T cell activation negative regulation of T cell proliferation T cell homeostasis negative regulation of sequence-specific DNA binding transcription factor activity negative regulation of interferon-gamma biosynthetic process negative regulation of interleukin-2 biosynthetic process negative regulation of transcription, DNA-templated positive regulation of transcription, DNA-templated positive regulation of transcription from RNA polymerase II promoter negative regulation of activated T cell proliferation negative regulation of isotype switching to IgE isotypes regulation of isotype switching to IgG isotypes negative regulation of cytokine secretion negative regulation of immune response T cell receptor signaling pathway negative regulation of T-helper 17 cell differentiation
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