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Primary Ab dilution-(1:200) in PBS containing 10% goat serum |
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Primary Ab dilution-(1:200) in PBS containing 10% goat serum |
Publication protocol
Cells were fixed with 4% paraformaldehyde at room temperature for 15 min. After washing with PBS three times, cells were permeabilized with 0.25% Triton X-100 in PBS for 5 min, and then incubated in a blocking buffer containing 10% goat serum, followed by incubation with anti-LC3 antibody (1:200) in PBS containing 10% goat serum overnight at 4°C. After incubating with 0.1% DAPI for 5 min at room temperature and another washing step with PBS, secondary Rhodanmine Red-X labelled antibody (1:100) was applied for 60 min. After washing with PBS, coverslips were transferred onto glass slides. Images were captured on a wide-field fluorescent microscopy (Zeiss).
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