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

Antibody Staining Kit

Supplier:  Invitrogen 72577540

Catalog No. 50-169-75

Includes: Fixation/Permeabilization Concentrate: 30mL (4X). Store at 2-8°C. Avoid agitation. Dilute 1 part Fixation/Permeabilization Concentrate with 3 parts Fixation/Permeabilization Diluent to make the Fixation/Permeabilization working solution. Caution: This solution contains Paraformaldehyde, which is toxic and a suspected carcinogen. Contact with eyes, skin and mucous membranes should be avoided. Wear proper protective clothing and gloves.
Fixation/Permeabilization Diluent: 100mL. Store at 2-8°C. The diluent is intended to be used in combination with the Fixation/Permeabilization Concentrate.
Permeabilization Buffer (10X): 100mL. Store at 2-8°C. Dilute to 1X with deionized/distilled water and store at 4°C. Caution: Harmful if swallowed or irritant by contact. Wear proper protective clothing and gloves.
anti-mouse/rat Foxp3 (FJK-16s): 25μg. Store at 2-8°C in the dark.

Description

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 49-55kDa 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)

Specifications

• Antigen: Foxp3
• Classification: Monoclonal
• Concentration: 0.2mg/mL
• Host Species: Rat
• Regulatory Status: RUO
• Target Species: Mouse, Rat
• Isotype: IgG2a κ

• Applications: Flow Cytometry
• Clone: FJK-16s
• Conjugate: PE
• Quantity: Kit
• Primary or Secondary: Primary
• Content And Storage: Store at 2-8°C. Light sensitive material. Use within 6 months of opening or by date indicated on the bottle.
• Product Line: eBioscience™

For Research Use Only. Not for use in diagnostic procedures.