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J. Biochem, 1970, Vol. 68, No. 1 119-124
© 1970 Japanese Biochemical Society

research-article

An Improved Method for the Determination of Tyrosine Residues in Proteins Using 1-Nitro-2-napthol

Kihachiro UEHARA, Shigeyasu MANNEN and Kenichi KISHIDA

The Laboratory of Biochemistry, Faculty of Pharmaceutical Sciences, Osaka University Toyonaka Osaka

A simple and reliable method for the quantitative determination of free tyrosine and. tyrosine residues in proteins is described. The method involves alkali-denaturation of proteins in a boiling water bath, the color forming reaction between tyrosine and 1-nitroso-2-naphthol in 19 N H2SO4, and measurement of the absorhance at 520 m µ. The method is preferable because of its simplicity, high sensitivity, reproducibility and remarkable stability of the red color. The procedure has been shown to be accurate with a wide variety of enzyme proteins.


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