Pyridylethylation for Quantification of Cysteine

Alkylation of cysteine with 4-vinylpyridine produces pyridylethylcysteine, which is stable to acid hydrolysis (Friedman et al., 1970) and readily measured by PTC-AAA. Pyridylethylcysteine (Pec) is commercially available (Pierce) and can be used to determine an appropriate Pec response factor as described for other standard amino acids. The following protocol is adapted from Hawke and Yuan (1987).

Additional Materials (also see Basic Protocol)

1:3 (v/v) 1 M TrisHCl (pH 8.5) containing 4 mM EDTA (Sigma)/8 M guanidineHCl or 1 M TrisHCl/1 mM EDTA, pH 8.5, containing 8 M urea (ultrapure)

1 M TrisHCl, pH 8.5, or 1 N HCl, for adjusting pH if necessary

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