Elsevier

Analytical Biochemistry

Volume 197, Issue 1, 15 August 1991, Pages 83-90
Analytical Biochemistry

A simplified formaldehyde fixation and immunoprecipitation technique for studying protein-DNA interactions

https://doi.org/10.1016/0003-2697(91)90359-2Get rights and content
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Abstract

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.

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Present address: Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, 250 Longwood Ave., Boston, MA 02115.