1. Fixation (PFA 4%, 10’, RT)
  2. Wash (PBS, 3 times, RT)
  3. Permeabilization (PBS+0,1%TRITON X-100+0,2%BSA, 10’, RT)
  4. Wash (PBS, 3 times, RT)
  5. Blocking (PBS+2%BSA, 30’, RT)
  6. Wash (PBS, 3 times, RT)
  7. Primary antibody incubation (dilute antibody in PBS+2%BSA, 60’, RT)
  8. Wash (PBS, 3 times, RT)
  9. Secondary antibody incubation (dilute antibody in PBS+2%BSA, 40’, RT, light protected)
  10. Wash (PBS, 3 times, RT)
  11. DAPI staining (dilute DAPI from stock solution in PBS at a final concentration of 0.1-10 μg/ml, 5’, RT, light protected)
  12. Wash (PBS, 3 times, RT)
  13. Wash (dH2O, 1 time)
  14. Mount