小鼠调节性T细胞检测试剂盒1

小鼠调节性T细胞检测试剂盒1
Mouse Regulatory T Cell Staining Kit #1
Also known as: Forkhead Box P3, FJK-16s PE; CD4 FITC, CD25 APC
RUO: For Research Use Only. Not for use in diagnostic procedures.
Description: The FJK-16s antibody reacts with mouse/rat 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 mouse Foxp3 staining sets and protocol reveals approximately 2% of total splenocytes 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. These data are consistent with a recent report which follows expression of Foxp3, using a GFP knock-in (Fontenot et al, 2005).

FJK-16s cross-reacts with rat Foxp3. This has been demonstrated by intracellular staining of Foxp3 and flow cytometry of rat splenocytes using the same method and reagents as used for mouse tissue. Please note that the CD4 and CD25 antibodies included in this kit only recognize the mouse antigens. For staining rat tissue, please use (CD4 FITC cat. 11-0400 and CD25 cat. 13-0390)

The anti-mouse Foxp3 Staining Set has been formulated and optimized for the staining of mouse splenocytes with the FJK-16s monoclonal antibody

Not included:
Isotype controls for anti-CD4 (rat IgG2a, cat. 11-4321) and anti-CD25 (rat IgG1, cat. 17-4301)

Reactivity	Mouse
Components	Anti-Mouse CD4 FITC (RM4-5): 50 µg. Use at 0.125 µg/test. Store at 2-8°C in the dark. Anti-Mouse CD25 APC (PC61.5): 50 µg. Use at 0.06 µg/test. Store at 2-8°C in the dark. Anti-Mouse/Rat Foxp3 PE (FJK-16s): 25 µg. Use at 0.5 µg/test. Store at 2-8°C in the dark. Rat IgG2a Isotype Control PE: 50 µg. Use at 0.5 µg/test. Store at 2-8°C in the dark. Anti-Mouse CD16/32 (Fc Block) Purified: 50 µg. Use at 1-5 µg/test. Store at 2-8°C. Flow Cytometry Staining Buffer: 200 ml. Store at 2-8°C. Fixation/Permeabilization Concentrate: 30 ml. Store at 2-8°C. Avoid agitation. This is a 4X stock solution that must be diluted prior to use with the Fixation/Permeabilization Diluent. Dilute 1 part Fixation/Permeabilization Concentrate with 3 parts Fixation/Permeabilization Diluent. Use within 6 months of receipt. 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: 100 ml. Store at 2-8°C. The diluent is intended to be used in combination with the Fixation/Permeabilization Concentrate. Permeabilization Buffer (10X): 100 ml. 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. Note: The 10X Permeabilization Buffer has a natural tendency to precipitate, however, its function is not affected by this. To clarify, the solution can be filtered after dilution to 1X working solution.
Reported Applications	Intracellular Staining Followed by Flow Cytometric Analysis 88-8115-40 Mouse Regulatory T Cell Staining Kit #3 Kit Kit eBioscience 5,388.00

88-8118-40

Mouse Regulatory T Cell Staining Kit #2 Kit

Kit

eBioscience

5,868.00