Sc46668

Cells seeded on chamber slides (Ibid, Munich, Germany) were washed with warm PBS and fixed with warm 4% paraformaldehyde (diluted in DMEM, 10% FCS) at 37°C for 15 min. For permeabilization, the cells were incubated 3-times with warm 0.1% Triton X-100/PBS for 5 min at 37°C and washed then 3-times with PBS for 10 min. The first antibodies (anti-fascin, Santa Cruz sc21746; anti VASP, Santa Cruz sc-46668) were diluted 1:200 in PBS/4% FCS and incubated over night at 4°C. After washing 3-times with PBS for 10 min, the secondary Alexa-fluor^®488 conjugated antibody (diluted 1:2000 in PBS/4% FCS) was added and the cells were incubated for 1 hour RT in the dark. Finally, the cells were washed 3-times with PBS and analyzed by fluorescence microscopy.