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Now it's time for the procedure itself

Those discussions typically mix opinions, anecdotes, and personal speculation, and they should not be confused with formal research or professional guidance

Subsequently, the cells were fixed with 4% paraformaldehyde at 16 C for 15 min, permeabilized in phosphate buffered saline (PBS) with 0.1% Triton X-100 for 5 min at 16 C, and incubated with first antibodies at 4 C overnight, followed by incubation with FITC-conjugated secondary antibodies at 16 C for 1 h

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