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hello today we will discuss about Western blot or protein immunoblot Western blotting or immunoblotting is an analytical technique is to detect a target protein in a sample by using cloning polyclonal or monoclonal antibody specific to that protein the mixture of proteins can be separated based on their size using polyacrylamide gel electrophoresis the protein bands are transferred to the blotting membrane the most widely used membrane for western blotting or nitrocellulose and polyvinyl in fluoride PVDF membranes so once the gentleman is completed general is carefully removed from the plate and soft in Western blot transfer buffer the question blow transfer buffer recipe includes Tris glycine and SDS and it has a pH of 8.4 generally he methanol is added to the transfer buffer letamp;#39;s see the membrane arrangement filter papers are arranged on the top and followed by gel then the Western blot membrane and a few filter papers at the bottom this is the arrangement you can see there