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After incubation, the medium was aspirated and cells were washed twice in PBS, fixed in 4% paraformaldehyde for 15-30 minutes, permeabilized in 0.2% Triton X-100 for 10 minutes at room temperature for the exposure of intracellular antigen, and blocked in PBS containing 3% bovine serum albumin (BSA
Step 2: Visual inspection Examine the solution for cloudiness, particles, color changes, or any deviation from its normal appearance
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