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Frederick Sanger’s protein sequencing method was a groundbreaking technique that allowed scientists to determine the exact amino acid sequence of proteins. This method, first developed in the 1950s, was primarily applied to insulin, and it laid the foundation for understanding protein structure and function. The method relies on the principle of selectively labeling the N-terminal amino acid of a protein and then identifying it after partial hydrolysis. The first step in Sanger’s method involves isolating the protein in a pure form. Once a homogeneous sample of the protein is obtained, it is treated with 1-fluoro-2,4-dinitrobenzene (FDNB), which reacts specifically with the free amino group at the N-terminal of the polypeptide chain. The reaction forms a dinitrophenyl (DNP) derivative of the N-terminal amino acid. _______ __________ _____ ______ ___ _____ _______ _______ _________ ________.
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