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in this video we will show you how to prepare a western blot membrane for chemiluminescent detection after antibody incubation and washing the membrane is ready to be incubated in enhanced chemiluminescent or ecl substrate prepare enough ecl substrate for the blot for a mini-sized membrane use about seven milliliters of substrate increase the volume to 12 milliliters if using a medium-sized membrane these volumes ensure uniform coverage and optimal results when too little substrate is used residual wash buffer is more likely to adversely affect the performance of the horseradish peroxidase enzyme although any small container will work for substrate incubation a clean sheet protector is recommended as it reduces the volume of substrate needed sheet protectors are also inexpensive and disposable incubate for five minutes to ensure the substrate has enough time to uniformly coat the membrane and displace the wash buffer membranes can also be kept inside a sheet protector for imagi
