标题：调节性T细胞（Treg）系列6－FOXP3的胞内染色步骤
- Perform cell surface staining as described in BioLegend’s Cell Surface Immunofuorescence Staining Protocol.
- 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.
- Wash once with cell staining bufer (Cat. No. 420201) by spinning at 250 X g for 5 minutes and remove the supernatant.
- Wash once with 1ml 1X BioLegend’s FOXP3 Perm bufer.
- Re-suspend cells in 1ml 1X BioLegend’s FOXP3 Perm bufer, 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 bufer.
- Add appropriate amount of fuorochrome conjugated anti-FOXP3 antibody and incubate at room temperature in the dark for 30 minutes.
- Wash twice with cell staining bufer, and resuspend in 0.5ml cell staining bufer then analyze with fow cytometer with appropriate instrument setting.
NOTE: BioLegend’s FOXP3 Fix/Perm Bufer Set (Cat. No. 421403) is specially developed and formulated for intracellular staining FOXP3 with minimum efect on surface fuorochrome staining and is highly recommended for optimal FOXP3 intracellular immunofuorescent staining results.
Related Information:
1. Roncador, G., et al., 2005 Eur. J. Immunol. 35:1681.
Reagent List:
1. Cell Staining Bufer (Cat. No. 420201)
2. FOXP3 Fix/Perm Bufer Set (Cat. No. 421403)
