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N-terminal Analysis of a Dipeptide

This Unilever Laboratory Experiment, published in 1967, describes a procedure based on the use of 1 -fluoro 2,4 dinitrophenylation (FDNB) as developed by Sanger during his studies on the structure of the protein hormone insulin. In the experiment, two dipeptides are used which contain the same two amino acids but in different sequence. Hence normal amino acid analysis cannot not differentiate them, but N-terminal analysis can. The suggested procedure is challenging and involves hazardous chemicals. After treatment with FDNB, hydrolysis in sealed tubes in an oven takes 5-6 hours. The resulting mixtures are analysed by paper chromatography.

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