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建立一种用 ＨＰＬＣ测定细胞中蛋白激酶活性的方法。方法：μＲｏｎｄｏｐｋａ Ｃ１８柱；流动相：甲醇：磷酸盐缓冲液（２０：８０ｖ／ｖ，内含５ ｍｍｏｌ／Ｌ ＰｉＣＡ，ｐＨ ７．０）；检测波长为２５９ ｎｍ；柱温：室温；灵敏度：０．０１ ＡＵＦＳ；进样 ２０ μｌ。结果：酶蛋白用量 ０． ３ ｍｇ左右、反应时间为 １０ ｍｉｎ、Ｈｉｓｔｏｎｅ（ Ⅲ－Ｓ）用量 １ｍｇ／ｍｌ、 ＡＴＰ浓度 ５００ μｍｏｌ／Ｌ时，活性测定结果理想。ＡＴＰ回收率为９９．６３％，ＲＳＤ为１．１１％（ｎ＝６）。结论：该方法简便、准确，适合于蛋白激酶的活性测定。
A method for the determination of protein kinase activity in cells by HPLC was established. Method: μRondopka C18 column; mobile phase: methanol: phosphate buffer (20:80 v / v, containing 5 mmol / L PiCA, pH 7.0); detection wavelength at 259 nm; .01 AUFS; Injection 20 μl. Results: The amount of enzyme protein 0. 3 mg, the reaction time was 10 min, the amount of Histone (Ⅲ-S) was 1 mg / ml and the concentration of ATP was 500 μmol / L. ATP recovery was 99.63% and RSD was 1.11% (n = 6). Conclusion: This method is simple and accurate and is suitable for the determination of protein kinase activity.