Imunocytochemistry on primary murine neural stem cell cultures Cells were washed with PBS once and fixed in 4%PFA/PBS for 10 min at RT. Cells were permeabilized with 0.2% trition X-100/PBS (1ml/each well) for 20 min at RT Cells were blocked using 10%BSA/0.1% trition X-100/PBS (1ml/each well) for 30 min at RT Ab42471 was added at 1:500 and incubated over night at 4?. Cells were washed twice with 0.1% trition X-100/PBS. Cells were incubated with secondary Ab (Flourescein anti-mouse) at RT for 1 hour in the dark (1:200), followed by being washed with 0.1% trition X-100/PBS twice A: DAPI staining of the nuclei B: SOX3 staining using Ab42471 Image submitted courtesy of: Shih-Han Lee
Anti-SOX3 antibody (ab42471) at 0.5 µg/ml + Raji cell lysate at 10 µgSecondaryHRP conjugated anti-Rabbit IgG at 1/50000 dilution
Performed under reducing conditions.