A SIMPLIFIED FORMALDEHYDE FIXATION AND IMMUNOPRECIPITATION TECHNIQUE FOR STUDYING PROTEIN DNA INTERACTIONS

被引：88

作者：

DEDON, PC

SOULTS, JA

ALLIS, CD

GOROVSKY, MA

机构：

[1] SYRACUSE UNIV, DEPT BIOL, BIOL RES LABS, SYRACUSE, NY 13244 USA

[2] UNIV ROCHESTER, DEPT BIOL, ROCHESTER, NY 14627 USA

来源：

ANALYTICAL BIOCHEMISTRY | 1991年 / 197卷 / 01期

基金：

美国国家卫生研究院;

关键词：

D O I：

10.1016/0003-2697(91)90359-2

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

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. © 1991.

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收藏

页码：83 / 90

页数：8

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