UCSF

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.