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[Music] hello everyone this video will discuss the immunodetection step of a western blot and well cover the purpose equipment and supplies and technique first off what is immunodetection immunodetection is used to detect specific proteins using polyclonal or monoclonal antibodies monoclonal antibodies bind to a single epitope within a target antigen conversely polyclonal antibodies consist of a mixed pool of immunoglobulin molecules that bind to several different epitopes found on a single antigen to perform immunodetection you will need a solution and buffer your primary and secondary antibody and substrate after blocking a primary antibody specific to the target protein is incubated with the membrane and the primary antibody binds to the target protein on this membrane after rinsing the membrane to remove unbound primary antibody the membrane is exposed to a specific enzyme conjugated secondary antibody the secondary antibody binds to the primary antibody which is reacted with the