FOXP3 Monoclonal Antibody (150D/E4), PE, eBioscience™

Product Specific Information

Description: eBioscience offers a panel of monoclonal antibodies to different epitopes of human/primate and mouse/rat Foxp3 protein, providing useful tools for investigating the complete expression pattern of Foxp3 at the protein level, and discerning the precise subsets of Foxp3+ cells. Other antibodies to Foxp3 available from eBioscience, which have been used significantly in scientific literature, include the anti-human Foxp3 PCH101 (Product # 72-5776) and ebio7979 (Product # 12-7979), and the anti-mouse/rat Foxp3 FJK-16s (Product # 72-5775). The 150D/E4 has been mapped to the splice region in Exon 2 found in human cells. The splicing of this region has not been shown to occur in mouse. Characterization of this antibody to the splice variant has been limited to epitope mapping. It should be noted that other antibodies such as PCH101 and 236A/E7 will recognize both the spliced and full length forms of the Foxp3 protein and stain more intensely than 150D/E4 and FJK-16s in human cells.

The 150D/E4 antibody reacts with mouse/rat/human 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.

Applications Reported: This 150D/E4 antibody has been reported for use in intracellular staining followed by flow cytometric analysis.

Applications Tested: This 150D/E4 antibody has been pre-titrated and 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 see BestProtocols® 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 5 µL (0.5 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.

Excitation: 488-561 nm; Emission: 578 nm; Laser: Blue Laser, Green Laser, Yellow-Green Laser.

Filtration: 0.2 µm post-manufacturing filtered.

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.