Anti-Mouse/Rat Foxp3 Staining Set APC, eBioscience™

The FJK-16s antibody reacts with mouse, rat, dog, porcine, and bovine 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.

Intracellular staining of mouse splenocytes with FJK-16s using the PE anti-mouse/rat Foxp3 Staining Set and protocol reveals approximately 2% of total cells in the C57Bl/6 strain and approximately 3-5% 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 FJK-16s. B220+, CD11b+, CD11c+, and Ly6G/Gr-1+ cells do not show significant co-staining with FJK-16s.

Please see our FAQ regarding the usage of eBioscience Foxp3 reagents.

Not included:
Fc Block (cat. 14-0161)
Flow Cytometry Staining Buffer (cat. 00-4222)
Rat IgG2a isotype Control (cat. 12-4321, 11-4321, or 17-4321)

Isotype
IgG2a, kappa

Laser
Red Laser

Emit
660 nm

Excite
633 - 647 nm

Reported Application
Intracellular Staining Followed by Flow Cytometric Analysis