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 ft.scurfy ft. (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 staining followed by flow cytometric analysis.Applications Tested: This FJK-16s antibody has been tested by intracellular staining with flow cytometric analysis of mouse splenocytes using the Foxp3/Transcription Factor Staining Buffer Set (cat. 00-5523) and protocol. Please refer to Best Protocols: Protocol B: One step protocol for (nuclear) intracellular proteins located under the Resources Tab online. This can 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. Alexa Fluor® 700 emits at 723 nm and can be excited with the red laser (633 nm). Most instruments will require a 685 LP mirror and 710/20 filter. Please make sure that your instrument is capable of detecting this fluorochrome. Excitation: 633-647 nm; Emission: 723 nm; Laser: Red Laser. Filtration: 0.2 µm post-manufacturing filtered. 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.
|Bovine, Canine, Feline, Mouse, Porcine, Rat|
|4° C, store in dark, DO NOT FREEZE!|
|Alexa Fluor® 700|
|PBS with 0.1% gelatin and 0.09% sodium azide; pH 7.2|
For Research Use Only.
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