体外激酶试验 |
体外HDAC实验步骤如下,用myc标记的HDAC1的质粒转染HeLa细胞,得到的免疫沉淀物先洗脱,用HD缓冲液(20 mM Tris-HCl, pH 8.0, 150 mM NaCl, 10% glycerol)重悬,分装冷冻储存。将HDAC1加入到含有40,000 cpm 3H标记的乙酰化组蛋白、 200 μlHD缓冲溶液、含有或不含有valproic acid的试管中。于37 °C旋转混匀2h,加入50 μl终止溶液终止反应,提取并分析(闪烁计数)带有3H标记的acetic acid。 |
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