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in a western blot the blocking and tagging portion will begin after the target protein has been transferred from the sds page into a pbdf membrane after the protein has been transferred into the membrane it will be incubated in a protein containing solution such as non-fat dry milk or purified protein like bovine serum albumin these proteins will cover any remaining area of the membrane which do not contain the target protein blocking the rest of the membrane will prevent any false signals and non-specific binding that could appear when imaging any excess blocking solution will be washed away following the wash the membrane is incubated in primary antibodies these primary antibodies are specific to the protein targeted and well bind to a his tag encoded in the protein after the incubation period is over any excess primary antibody will be washed off washing away the excess will prevent non-specific binding signals following the wash the membrane is incubated with secondary antibodies t