Centrifuge fixed cells and remove supernatant. Add Cell Permeabilization Buffer directly to fixed cells, using approximately 100 µl per 1 million cells. Incubate for 10 min at room temperature. Cells may be washed with phosphate-buffered saline (PBS) to remove Cell Permeabilization Buffer prior to antibody incubation, but it is not necessary. Note that the Triton™ X-100 detergent can reduce cell pellet adhesion, so care should be taken to retain cells when removing supernatant following centrifugation.
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