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hello everyone this is Sean Taylor field application scientist manager for bio-rad in Canada and in this video presentation Iamp;#39;ll be discussing the quantitative Western blotting workflow and how optimization is critical for achieving precise accurate and reproducible data the Western blotting workflow involves four key steps the first of which is the separation of protein lysates on a gel by their molecular weight the second step involves the transfer of the gel separated proteins to a membrane using a transfer apparatus the third step which is in of itself a multi-step process involves the incubation of the protein transferred membrane with the primary and secondary antibodies and the final step involves the revelation of the membrane in a fluorescence and/or chemiluminescence enabled imaging system of course the mode of detection in the imaging system depends on the secondary antibody and whether or not itamp;#39;s conjugated to horseradish peroxidase for chemilumines
