Thermo Fisher Scientific FOXP3 Monoclonal Antibody (FJK-16s), PerCP-eFluor 710, eBioscience

Applications

Tested Dilution

Publications

Western Blot (WB)

-

View 2 publications 2 publications

Immunohistochemistry (IHC)

-

View 17 publications 17 publications

Immunocytochemistry (ICC/IF)

-

View 2 publications 2 publications

Flow Cytometry (Flow)

0.25 µg/test

View 92 publications 92 publications

Product Specifications

Species Reactivity

Bovine, Dog, Cat, Mouse, Pig, Rat

Published species

Human, Mouse

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 PerCP-eFluor™ 710 PerCP-eFluor™ 710

View additional formats

• Unconjugated
• Alexa Fluor 488
• Alexa Fluor 532
• Alexa Fluor 561
• Alexa Fluor 660
• Alexa Fluor 700
• APC
• Biotin
• Brilliant Violet 421
• Brilliant Violet 605
• Brilliant Violet 711
• Brilliant Violet 786
• 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
• Request custom conjugation

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!

Shipping conditions

Ambient (domestic); Wet ice (international)

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 (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 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.