Electrophoretic separation of alkaline and acid phosphatase isoenzymes from the pulp of monkey teeth.
Electrophoretic separation of alkaline and acid phosphatase isoenzymes from the pulp of monkey teeth.
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从猴牙髓中电泳分离碱性和酸性磷酸酶同工酶。
DOI：
10.3109/00016357809029090
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发表时间：
1978
影响因子：
2
通讯作者：
G. Hasselgren
中科院分区：
文献类型：
--
作者：
A. Franzén;G. Hasselgren
将猴浆在 Triton tris 溶液中均质化。三个离心步骤（800、20000 和 105000 g）后，将上清液涂在 pH 7.5 的三-二乙基巴比妥酸缓冲液中的丙烯酰胺柱上。在 18--20 摄氏度下以每个凝胶柱 2.5 mA 的恒定电流进行电泳。使用萘酚-AS-MX-磷酸盐作为底物和固红紫 LB 盐作为偶联剂，在 pH 8.3 下进行碱性磷酸酶 (E.C. 3.1.3.1) 的孵育。酸性磷酸酶 (E.C. 3.1.3.2) 的孵育在 pH 5.0 下进行，使用 α-萘基磷酸酯作为底物，六氮化副玫瑰苯胺作为偶联剂。碱性磷酸酶和酸性磷酸酶孵育后，显示出两条显示酶活性的条带。通过在温育前进行加热（56℃）处理或向温育介质中添加钒酸盐或焦磷酸盐，表明快速移动的碱性磷酸酶带的主要部分对这些程序敏感。缓慢移动带的碱性磷酸酶似乎耐热或添加抑制剂。
Monkey pulps were homogenized in a Triton tris solution. After three centrifugation steps (800, 20000, and 105000 g) the supernatant was applied on acryl amide columns at pH 7.5 in a tris-diethyl barbituric acid buffer. Electrophoresis was performed at a constant current of 2.5 mA per gel column at 18--20 degrees C. Incubations for alkaline phosphatase (E.C. 3.1.3.1) were carried out at pH 8.3 using naphthol-AS-MX-phosphate as substrate and Fast Red Violet LB salt as coupler. Incubations for acid phosphatase (E.C. 3.1.3.2) were undertaken at pH 5.0 using alpha-naphtyl phosphate as substrate and hexazotized pararosanilin as coupling agent. After the incubations for alkaline phosphatase as well as acid phosphatase two bands showing enzyme activity were demonstrated. By means of treatment with heat (56 degrees C) prior to incubation or addition of vanadate or pyrophosphate to the incubation medium it was shown that the main part of the fast moving alkaline phosphatase band was sensitive to these procedures. The alkaline phosphatase of the slow moving band appeared to be resistant to heat or the addition of inhibitors.