Chromatin Immunoprecipitation of Low Number of FACS-Purified Epidermal Cells

Methods Mol Biol. 2020:2154:197-215. doi: 10.1007/978-1-0716-0648-3_17.

Abstract

Chromatin immunoprecipitation followed by sequencing (ChIP-seq) is a method designed to detect interactions between chromatin and the proteins bound to it. This method has been widely used for characterizing epigenetic landscapes in many cell types; however, a limiting factor has been the requirement of a high number of cells. Here, we describe a protocol for ChIP in epidermal cells from a newborn mouse, purified by fluorescence-activated cell sorting (FACS). This protocol has been optimized specifically for prefixed, low cell numbers, resulting in enough immunoprecipitated DNA suitable for genome-wide analysis.

Keywords: ChIP-seq; Epidermal cells; FACS purified cells; Low cell number ChIP.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Animals, Newborn
  • Blotting, Western
  • Chromatin Immunoprecipitation* / methods
  • Epidermal Cells / metabolism*
  • Flow Cytometry* / methods
  • Immunohistochemistry
  • Mice
  • Skin