Surface staining of BALB/c splenocytes with Anti-Mouse CD4 PE (Product # 12-0042) and Anti-Mouse CD25 APC (Product # 17-0251), and subsequently fixed and permeabilized using the Foxp3 Staining Buffers (Product # 00-5523) and stained with 1.0 µg of Rat IgG2a K Isotype Control FITC (Product # 11-4321) or 1.0 µg of Anti-Mouse/Rat Foxp3 FITC. The histogram (left) demonstrates Foxp3 staining (filled histogram) or isotype control staining (open histogram) after gating on CD4+CD25+ cells. The dot plot (right) demonstrates co-staining of CD4 and Foxp3. Cells in the lymphocyte gate were used for analysis.
|Tested species reactivity||Bovine, Dog, Cat, Mouse, Pig, Rat|
|Published species reactivity||Not Applicable|
|Host / Isotype||Rat / IgG2a, kappa|
|Storage buffer||PBS, pH 7.2, with 0.1% gelatin|
|Contains||0.09% sodium azide|
|Storage Conditions||4° C, store in dark, DO NOT FREEZE!|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||1 µg/test|
|Immunohistochemistry (Frozen) (IHC (F))||Assay-Dependent|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
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 flow cytometric analysis. For optimal staining with FJK-16s, it is highly recommended to use the Anti-Mouse/Rat Foxp3 FITC Staining Set (cat. 71-5775). FJK-16s has also been reported for use in immunohistochemical staining of frozen tissue sections.
Applications Tested: This FJK-16s antibody has been tested by intracellular flow cytometric analysis of mouse splenocytes using the Anti-Mouse Foxp3 FITC Staining Set (cat. 71-5775) and protocol. Please refer to intracellular Staining Protocol on Best Protocol web page - Protocol B: One-step protocol for intracellular (nuclear) proteins. This antibody can be used at less than or equal to 1 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.
Excitation: 488 nm; Emission: 520 nm; Laser: Blue Laser
FOXP3 is a member of the forkhead/winged-helix family of transcriptional regulators and is highly conserved across mammals. FOXP3 is essential for normal immune homeostasis. FOXP3 is 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 than other T cells.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
|Not Applicable||Not Cited||Sensitive detection of Foxp3 expression in bovine lymphocytes by flow cytometry.||Gerner W,Stadler M,Hammer SE,Klein D,Saalmüller A||Veterinary immunology and immunopathology (138:154)||2010|
||Detection of Foxp3 protein expression in porcine T lymphocytes.||Käser T,Gerner W,Hammer SE,Patzl M,Saalmüller A||Veterinary immunology and immunopathology (125:92)||2008|
||Use of FoxP3 expression to identify regulatory T cells in healthy dogs and dogs with cancer.||Biller BJ,Elmslie RE,Burnett RC,Avery AC,Dow SW||Veterinary immunology and immunopathology (116:69)||2007|
|Not Applicable||Not Cited||Detection of Foxp3 protein expression in porcine T lymphocytes.||Käser T,Gerner W,Hammer SE,Patzl M,Saalmüller A||Veterinary immunology and immunopathology (125:92)||2008|
|Not Applicable||Not Cited||Use of FoxP3 expression to identify regulatory T cells in healthy dogs and dogs with cancer.||Biller BJ,Elmslie RE,Burnett RC,Avery AC,Dow SW||Veterinary immunology and immunopathology (116:69)||2007|
|Not Applicable||Not Cited||A link between PDL1 and T regulatory cells in fetomaternal tolerance.||Habicht A,Dada S,Jurewicz M,Fife BT,Yagita H,Azuma M,Sayegh MH,Guleria I||Journal of immunology (Baltimore, Md. : 1950) (179:5211)||2007|
|Not Applicable||Not Cited||Shift towards pro-inflammatory intestinal bacteria aggravates acute murine colitis via Toll-like receptors 2 and 4.||Heimesaat MM,Fischer A,Siegmund B,Kupz A,Niebergall J,Fuchs D,Jahn HK,Freudenberg M,Loddenkemper C,Batra A,Lehr HA,Liesenfeld O,Blaut M,Göbel UB,Schumann RR,Bereswill S||PloS one (2:null)||2007|
|Not Applicable||Not Cited||Foxp3-expressing CD103+ regulatory T cells accumulate in dendritic cell aggregates of the colonic mucosa in murine transfer colitis.||Leithäuser F,Meinhardt-Krajina T,Fink K,Wotschke B,Möller P,Reimann J||The American journal of pathology (168:1898)||2006|