The antibody was purified by affinity chromatography, and conjugated with Pacific Blue™ under optimal conditions. The solution is free of unconjugated Pacific Blue™.
Concentration:
0.5 mg/ml
Storage & Handling:
The antibody solution should be stored undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
Application:
ICFC
Recommended Usage:
Each lot of this antibody is quality control tested by immunofluorescent staining for flow cytometric analysis, the suggested use of this reagent is ≤ 0.25 µg per 106 cells in 100 µl volume. It is highly recommended that the reagent be titrated for optimal performance for each application.
* Pacific Blue™ has a maximum emission of 455 nm when it is excited at 405 nm. Prior to using Pacific Blue™ conjugate for flow cytometric analysis, please verify your flow cytometer's capability of exciting and detecting the fluorochrome. ** Pacific Blue™ is a registered trademark of Molecular Probes, Inc. Pacific Blue™ dye antibody conjugates are sold under license from Molecular Probes, Inc. for research use only, except for use in combination with microarrays and high content screening, and are covered by pending and issued patents.
Application Notes:
FOXP3 Intracellular Staining Procedures: 1. Perform cell surface staining as described in BioLegend's Cell Surface Immunofluorescence Staining Protocol. 2. Add 1 ml of 1X BioLegend's FOXP3 Fix/Perm solution to each tube, vortex and incubate at room temperature in the dark for 20 minutes, then spin down the cells and remove the supernatant. 3. Wash once with cell staining buffer (Cat. No. 420201) by spin at 250Xg for 5 minutes and remove the supernatant. 4. Wash once with 1ml 1X BioLegend's FOXP3 Perm buffer. 5. Re-suspend cells in 1ml 1X BioLegend's FOXP3 Perm buffer, incubate at room temperature in the dark for 15 minutes, spin down cells and discard the supernatant, then resuspend the pellet in 100 µl of 1X BioLegend's FOXP3 Perm buffer. 6. Add appropriate amount of flurochrome conjugated anti-FOXP3 antibody and incubate at room temperature in the dark for 30 minutes. 7. Wash twice with cell staining buffer, and resuspend in 0.5ml cell staining buffer then analyze with flow cytometer with appropriate instrument setting. NOTE: BioLegend's FOXP3 Fix/Perm buffer set (Cat. No. 421403) is specially developed and formulated for intracellular staining FOXP3 with minimum effect on surface fluorochrome staining and is highly recommended for optimal result of FOXP3 intracellular immunofluorescence staining.
Application References:
1. Ono,M., et al: Nature 2007 446:685
C57BL/6 splenocytes surface stained with CD4 (GK1.5) PE and then intracellularly stained with MF-14 Pacific Blue™
Description:
FOXP3 is a transcription factor of 50 kd. It is a master regulatory gene and specific marker of T reg cells.Originally identified as the defective gene in Scurfy mice which spontaneously develop autoimmune diseases.It is now reported that FoxP3 is mainly expressed in CD4+CD25+ T reg cells as a specific marker for T reg.
Other Names:
Forkhead box protein P3, Scurfin, JM2, IPEX, Zinc finger protein JM2
