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ChIP is an antibody-based technique to enrich DNA-binding proteins and their targets. Learn about native and crosslinked ChIP, ChIP-PCR, ChIP-chip, and ChIP-seq methods and how they are used to study protein-DNA interactions.
Chromatin immunoprecipitation ( ChIP) is a type of immunoprecipitation experimental technique used to investigate the interaction between proteins and DNA in the cell.
What is a ChIP Assay? Chromatin immunoprecipitation (ChIP) assays are used to evaluate transcription factor-DNA interactions and are critical for advancing gene expression regulation and epigenetic modifications studies. ChIP can detect and relatively quantify specific protein-DNA and protein-protein interactions in vivo at a single locus or ...
- Crosslinking. ChIP assays begin with covalent stabilization of the protein–DNA complexes. Many protein–DNA interactions are transient and involve multiprotein complexes to orchestrate biological functions.
- Cell lysis. In this step, cell membranes are dissolved with detergent-based lysis solutions to liberate cellular components, and crosslinked protein–DNA complexes are solubilized.
- Chromatin preparation (shearing/digestion) The extraction step yields all nuclear material, which includes unbound nuclear proteins, full-length chromatin, and the crosslinked protein–DNA complexes.
- Immunoprecipitation. To isolate a specifically modified histone, transcription factor, or cofactor of interest, ChIP-validated antibodies are used to immunoprecipitate and isolate the target from other nuclear components.
One of the widely used assays to study this is chromatin immunoprecipitation (ChIP) assay, which enables analysis of association of regulatory molecules to specific promoters and histone modifications in vivo.
- Padmaja Gade, Dhan V. Kalvakolanu
- 10.1007/978-1-61779-376-9_6
- 2012
- 2012
Learn how to perform chromatin immunoprecipitation (ChIP) assays to study protein–DNA interactions and epigenetics. Find out the steps, methods, reagents and products for successful ChIP experiments.
Learn how to perform the chromatin immunoprecipitation (ChIP) assay to probe protein-DNA interactions in the cell. The web page provides an overview of the ChIP assay, a kit for cells and tissues, and references for further reading.
