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Protein sequencing
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=== Reaction === The peptide to be sequenced is [[adsorption|adsorbed]] onto a solid surface. One common [[Substrate (biochemistry)|substrate]] is glass fibre coated with [[polybrene]], a [[cationic polymer]]. The Edman reagent, [[phenylisothiocyanate]] (PITC), is added to the adsorbed peptide, together with a mildly basic [[buffer solution]] of 12% [[trimethylamine]]. This reacts with the amine group of the N-terminal amino acid. The terminal amino acid can then be selectively detached by the addition of [[anhydrous]] acid. The derivative then [[isomerization|isomerises]] to give a substituted [[phenylthiohydantoin]], which can be washed off and identified by chromatography, and the cycle can be repeated. The efficiency of each step is about 98%, which allows about 50 amino acids to be reliably determined. [[File:Protein sequencer.jpg|A Beckman-Coulter Porton LF3000G protein sequencing machine|thumb]]
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