The term "Western blotting" is commonly used in molecular biology to describe a technique for detecting specific proteins from a sample. The word "Western" is pronounced /ˈwɛstərn/ and refers to the direction in which the technique was developed - from the previous technique of Southern blotting. The word "blotting" is pronounced /ˈblɒtɪŋ/ and refers to the process of transferring proteins from a gel to a membrane. Overall, the spelling of "Western blotting" accurately reflects the sounds of the words when pronounced using IPA phonetic transcription.
Western blotting, also known as immunoblotting, is a molecular biology technique used to detect specific proteins from a mixture of proteins. It is a powerful tool used to study protein expression and to identify and analyze protein interactions. The technique is named after its similarity to the Southern blotting and Northern blotting techniques that are used to detect DNA and RNA, respectively.
In Western blotting, proteins are separated based on their molecular weight using gel electrophoresis. The protein mixture is first denatured and subjected to gel electrophoresis, which separates the proteins according to their size. After electrophoresis, the proteins are transferred from the gel onto a solid membrane, usually made of nitrocellulose or polyvinylidene difluoride (PVDF). This transfer process, known as blotting, preserves the spatial arrangement of the separated proteins.
Once the proteins are transferred onto the membrane, the membrane is then blocked to prevent non-specific binding. This is followed by incubation with a primary antibody that specifically recognizes the protein of interest. The primary antibody binds to the target protein, forming an antibody-antigen complex. The membrane is then washed to remove unbound primary antibody, and a secondary antibody is added. The secondary antibody is tagged with a reporter molecule, such as an enzyme or a fluorescent dye. Binding of the secondary antibody to the primary antibody-antigen complex allows for detection of the target protein using various detection methods, such as chemiluminescence or fluorescence.
Western blotting is a widely used technique in molecular biology research and diagnostic applications. It provides valuable information about protein expression, size, and interactions, allowing researchers to study various aspects of protein biology.
The term "Western blotting" was coined by scientists in 1979 as a variation of the well-established laboratory technique known as "Southern blotting". The technique was named after its inventor, Edwin M. Southern, who developed it in the 1970s.
Southern blotting was used to study DNA fragments and their characteristics by separating them through gel electrophoresis and then transferring them to a solid support membrane for analysis. This method was named after Southern because he was the first to describe it in a landmark scientific publication.
Later, in 1979, researchers in a laboratory at Stanford University led by George Stark decided to use a similar technique to study RNA instead of DNA. Since they were performing the technique in a lab situated in the western part of the United States (California), they called this RNA analysis technique "Western blotting" as a humorous nod to Southern blotting.