Immunoblotting, also known as Western blotting, is a widely used analytical technique in molecular biology and immunology to detect specific proteins in a sample of tissue homogenate or extract. It involves separating proteins by gel electrophoresis, transferring them to a membrane (typically nitrocellulose or PVDF), and then probing the membrane with specific antibodies that bind to the target protein. The bound antibodies are then detected, often using enzyme-linked secondary antibodies or fluorescently labeled antibodies, allowing for the identification and quantification of the protein of interest. Immunoblotting is commonly used to confirm protein expression, determine protein size, and assess post-translational modifications.
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