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