Biomedical Engineering Reference
In-Depth Information
Fig. 2
Flow chart to monitor cargo traffi cking from the plasma membrane to
intracellular compartments by pulse shape analysis (PulSA). Shown here is the
protocol for monitoring cargo traffi cking from the cell membrane to intracellular
compartments by PulSA. Briefl y, cells are detached from the plates or dishes
before incubation with epitope-specifi c antibodies for 30 min on ice. After
30 min, cells are washed in cold PBS before incubation with serum-free media
for different durations at 37 °C or fi xed (0 min). Cells were fi xed in 4 % PFA/PBS
for 15 min after each time points. Cells were then permeabilized in 0.1 % Triton
X-100 for 4 min and then blocked in 5 % FCS/PBS. After which, the cells can be
stained with additional organelle markers before fl ow cytometry
