Flow Cytometry Product Notes: Test Sizes: To accommodate multicolor flow cytometry, eBioscience is in the process of reducing test size volumes from 20 µl to 5 µl. Please check your antibody vial for the recommended test size. Fluorochrome Replacements: eBioscience is in the process of replacing all Alexa Fluor® 647 conjugated products with eFluor® 660 conjugated products.
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 (cat. 72-5776) and ebio7979 (cat. 12-7979), and the anti-mouse/rat Foxp3 FJK-16s (cat. 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
For research use only, not for diagnostic or therapeutic use. This 150D/E4 antibody has been reported for use in immunoblotting (WB), and immunohistochemical staining.
Applications Tested
This 150D/E4 antibody has been tested by immunoblotting of recombinant mouse and human Foxp3 protein.
The purified format is not recommended for intracellular staining and flow cytometric analysis.
References
Fontenot, JD., Rasmussen, JP., Williams, LM., Dooley, JL., Farr, AG., Rudensky AY. 2005. Regulatory T cell lineage specification by the forkhead transcription factor foxp3. Immunity. 22(3): 329-41.
Hori ,S., Nomura, T., Sakaguchi, S. 2003. Control of regulatory T cell development by the transcription factor Foxp3. Science. 299(5609):1057-61.
Roncador G, Brown PJ, Maestre L, Hue S, Martinez-Torrecuadrada JL, Ling KL, Pratap S, Toms C, Fox BC, Cerundolo V, Powrie F, Banham AH. Analysis of FOXP3 protein expression in human CD4+CD25+ regulatory T cells at the single-cell level.
Eur J Immunol. 2005 Jun;35(6):1681-91 (150D, IHC, PubMed).
Sakaguchi, S. 2003. The origin of FOXP3-expressing CD4+ regulatory T cells: thymus or periphery. J Clin Invest. 112(9):1310-2.
