IF Staining Cell Line

Immunofluorescence

1. Grow cell in wells.
2. Wash with PBS TWICE (500ul/well) .
3. Fix with 200ul of 4% paraformaldehyde/PBS for 20 min at RT (fix time)
   (Stock is 16%).
4. Permeabilize with 0.3% Triton x-100/PBS for 15 min at RT (fix time).
5. Treated with goat serum/PBS (1:200) for 30 min with shaker ~21 at RT.
6. Added anti-beta-catenin Ab in goat serum/PBS for 1.5 hr at RT (fix time)
7. Wash with PBS three times for 5 min each with shaker ~21 at RT
8. Added 100ul goat anti-mouse FITC(green, 1:500) for 45 min at RT. (fix time)
9. Wash with PBS three times for 10 min each at RT.