Description: The NRRF-30 antibody reacts with mouse 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 NRRF-30 antibody has mapped the epitope to amino acids 1-75 of the mouse Foxp3 protein. Intracellular staining of mouse splenocytes with fluorochrome-conjugated NRRF-30 using the eBioscience Foxp3 Staining Buffers (cat. 00-5523) and corresponding staining protocol reveals approximately 3% of total cells in the C57Bl/6 strain and approximately 5% in the BALB/c mouse strain. Multicolor flow cytometric analysis demonstrates approximately 90% of the CD4+/CD25+ cells and 4% of the CD4+/CD25- cells staining with NRRF-30. Co-staining with FJK-16s (anti-mouse/rat Foxp3 cat. 71-5775), which has been mapped to amino acids 71-125, and NRRF-30 shows 100% correlation, indicating that the same cells are stained with both anti-mouse Foxp3 antibodies. Please see our FAQ regarding the usage of eBioscience Foxp3 reagents.Applications Reported: This NRRF-30 antibody has been reported for use in intracellular staining followed by flow cytometric analysis. Applications Tested: This NRRF-30 antibody has been tested by intracellular flow cytometric analysis of mouse splenocytes using the Foxp3/Transcription Factor Staining Buffer Set (cat. 00-5523) and protocol. Please see Best Protocols Section (Staining Intracellular Antigens for Flow Cytometry) for staining protocol (refer to Protocol B: One-step protocol for intracellular (nuclear) proteins). 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. Excitation: 488-561 nm; Emission: 578 nm; Laser: Blue Laser, Green Laser, Yellow-Green 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.
|4° C, store in dark, DO NOT FREEZE!|
|PBS with 0.1% gelatin and 0.09% sodium azide; pH 7.2|
For Research Use Only.
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