Detergent Lysis of Tissue Culture Cells for Immunoprecipitation

Abstract

This protocol describes the lysis of tissue culture cells for the solubilization of proteins of interest for immunoprecipitation. Upon collection of cells by centrifugation and depending on the use of either Tris- or phosphate-based cell lysis buffers, cells are rinsed, respectively, in either TBS or PBS before lysis. If possible, the pH of the Wash buffer should match that of the Lysis buffer. Adherent cells can be directly lysed on the plate. This is particularly useful upon lysis encompassing mild nonionic detergents, leaving the cytoskeleton intact. Alternatively, adherent cells can be scraped off the plate and directly resuspended in Lysis buffer or Wash buffer for transfer to a tube followed by the addition of Lysis buffer.