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In this video tutorial, learn how to prepare a western blot membrane for chemiluminescent detection. After antibody incubation and washing, the membrane is ready for enhanced chemiluminescent (ECL) substrate. Use 7ml of substrate for a mini-sized membrane and 12ml for a medium-sized one. Use a clean sheet protector for substrate incubation to reduce volume needed. Incubate for five minutes to ensure uniform coating and displacement of wash buffer. Sheet protectors are recommended for easy disposal and storage of membranes.