Invitrogen

FOXP3 Monoclonal Antibody (FJK-16s), Brilliant Violet™ 786, eBioscience™

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Datasheet
Protocols
Questions & Answers
Tech Support

Product Details

417-5773-82

Applications
Tested Dilution
Publications

Flow Cytometry (Flow)

1.0 µg/test
-
Product Specifications

Species Reactivity

Bovine, Dog, Cat, Mouse, Pig, Rat

Host/Isotype

Rat / IgG2a, kappa

Recommended Isotype Control

Rat IgG2a kappa Isotype Control (eBR2a), Brilliant Violet™ 786, eBioscience™

Class

Monoclonal

Type

Antibody

Clone

FJK-16s

Conjugate

Brilliant Violet™ 786 Brilliant Violet™ 786 Brilliant Violet™ 786

Excitation/Emission Max

406/788 nm View spectra spectra

Form

Liquid

Concentration

0.2 mg/mL

Purification

Affinity chromatography

Storage buffer

PBS, pH 7.2, with BSA

Contains

0.09% sodium azide

Storage conditions

4° C, store in dark, DO NOT FREEZE!

Shipping conditions

Wet ice

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 (Product # 00-5523-00). 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 followed by 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 Flow Protocols. This may be used at less than or equal to 1.0 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.

Brilliant Violet™ 786 (BV786) is a tandem dye that emits at 786 nm and is intended for use on cytometers equipped with a violet (405 nm) laser. Please make sure that your instrument is capable of detecting this fluorochrome.

When using two or more Super Bright, Brilliant Violet™, Brilliant Ultra Violet™, or other polymer dye-conjugated antibodies in a staining panel, it is recommended to use Super Bright Complete Staining Buffer (Product # SB-4401-42) or Brilliant Stain Buffer™ (Product # 00-4409-75) to minimize any non-specific polymer interactions. Please refer to the datasheet for Super Bright Staining Buffer or Brilliant Stain Buffer for more information.

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 # 00-5333-54) 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.

Our internal testing suggests that Brilliant Violet™ 786 (BV786) is not compatible with methanol-based fixation.

Excitation: 407 nm; Emission: 786 nm; Laser: Violet Laser.

BRILLIANT VIOLET™ is a trademark or registered trademark of Becton, Dickinson and Company or its affiliates, and is used under license. Powered by Sirigen.™

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; forkhead/winged helix transcription factor 3; Foxp3; MGC141961; MGC141963; RP23-54C14.1; Scurfin; scurfy

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

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UniProt ID: (Mouse) Q99JB6

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

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Function(s)
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
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 differentiation CD4-positive, CD25-positive, alpha-beta regulatory T cell lineage commitment T cell mediated immunity tolerance induction tolerance induction to self antigen regulation of immunoglobulin production positive regulation of T cell tolerance induction 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 transcription, DNA-templated regulation of transcription, DNA-templated regulation of transcription from RNA polymerase II promoter transcription from RNA polymerase II promoter negative regulation of cell proliferation anatomical structure morphogenesis positive regulation of gene expression 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 negative regulation of sequence-specific DNA binding transcription factor activity regulatory T cell differentiation negative regulation of interferon-gamma biosynthetic process negative regulation of interleukin-2 biosynthetic process positive regulation of regulatory T cell differentiation negative regulation of transcription, DNA-templated positive regulation of transcription, DNA-templated positive regulation of transcription from RNA polymerase II promoter negative regulation of isotype switching to IgE isotypes regulation of isotype switching to IgG isotypes negative regulation of lymphocyte proliferation negative regulation of cytokine secretion negative regulation of inflammatory response negative regulation of immune response T cell receptor signaling pathway negative regulation of T-helper 17 cell differentiation biological_process response to virus
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