A SIMPLIFIED FORMALDEHYDE FIXATION AND IMMUNOPRECIPITATION TECHNIQUE FOR STUDYING PROTEIN DNA INTERACTIONS
A SIMPLIFIED FORMALDEHYDE FIXATION AND IMMUNOPRECIPITATION TECHNIQUE FOR STUDYING PROTEIN DNA INTERACTIONS
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DOI：
10.1016/0003-2697(91)90359-2
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发表时间：
1991-08-15
影响因子：
2.9
通讯作者：
GOROVSKY, MA
中科院分区：
文献类型：
--
作者：
DEDON, PC;SOULTS, JA;GOROVSKY, MA
利用单细胞真核生物嗜热四膜虫，开发了一种简单的方法，用于研究蛋白质-DNA协会通过交联蛋白质与DNA与甲醛和免疫沉淀的溶解的染色质片段与特异性抗血清。该方案使用粗抗血清，只涉及三个步骤：交联，剪切溶解染色质，和免疫沉淀。某些关键参数，如固定时间和NaCl浓度的优化方法进行了说明。该方法可能对各种核抗原普遍有用。
Using the single cell eukaryote Tetrahymena thermophila, a simple method was developed for studying protein-DNA associations by cross-linking proteins to DNA with formaldehyde and immunoprecipitating the solubilized chromatin fragments with a specific antiserum. The protocol uses crude antiserum and involves only three steps: cross-linking, shearing to solubilize the chromatin, and immunoprecipitation. Methods for optimizing certain critical parameters, such as fixation time and NaCl concentration, are described. The method is likely to be generally useful for a variety of nuclear antigens.