When this antibody binds to the protein on the membrane, it can be detected with a chemiluminescent or fluorescent tagged secondary antibody allowing for visualization of the protein band. This step involves incubating the transfer membrane in a solution containing an antibody to the protein of interest. This is the procedure that I will highlight in this article. Getting the separated proteins out of the gel and bound to a membrane allows for easier detection. Separation of the proteins in your lysate by molecular weight is done through electrophoresis. Read our lysate preparation protocol here. You can read about our recommended lysis buffer here. You will never be able to visualize your proteins if they remain trapped in your tissue or cells. The number one thing you can do for success in your Western blot is appropriate sample prep. What are the steps of a Western blot procedure? Two years after that, when Towbin transferred proteins to a membrane, it eventually became known as a “Western blot”. Two years later, researchers at Stanford developed a similar RNA detection technique that was dubbed “Northern blot”. It involved transferring DNA to a membrane and was dubbed a “Southern blot” in honor of its inventor’s name. Edwin Southern published a DNA detection technique in 1975 from his lab in England. “Western” is a play on words based on a similar technique. It was introduced in 1979 by Harry Towbin’s research lab in Switzerland. The Western Blot (or immunoblot) technique uses antibodies to detect protein targets that have been bound to a membrane. The Origin and Methods of Western Blotting What is Western Blotting?
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