chromatin immunoprecipitation

Editor’s Note: At the author’s request on August 20th, 2015, revisions to this article were made following careful consideration. All changes are denoted in red. In the previous post of this ChIP series, we left off with your samples rotating at four degrees overnight, to give the antibodies plenty of time to bind and the beads plenty of time to block.  In this final post, we will finish the assay and provide advice on how to QC and analyze your libraries. [more…]

Editor’s Note: At the author’s request on August 20th, 2015, revisions to this article were made following careful consideration. All changes are denoted in red. In the first post of this ChIP series, we introduced the reader to the general concept and workflow of chromatin immunoprecipitation (ChIP), and provided advice on planning a ChIP experiment and preparing the necessary reagents.  In this post, we will go through a more detailed protocol that we have used in our lab with success that [more…]

Editor’s Note: At the author’s request on August 20th, 2015, revisions to this article were made following careful consideration. All changes are denoted in red. Since its introduction in the 1980s, chromatin immunoprecipitation (ChIP) has become one of the most important and powerful techniques in the field of genetics, allowing researchers to characterize a given protein’s binding sites across the genome.  With the advent of next-generation sequencing and whole genome analysis, a single ChIP can deliver gigabytes of information, including not [more…]

Every epigenetics scientist knows that chromatin immunoprecipitation (ChIP) is a valuable technique for studying protein-DNA interactions.  They also know that antibodies used in ChIP to capture the DNA/protein complex must be reliable and specifically recognize the fixed protein that is bound to the chromatin complex. But how can a researcher be certain that the antibodies they are using work well in ChIP?  For many researchers it is not always the case to be “blessed” with a quality ChIP-grade antibody and, unfortunately, [more…]

Researchers rely on high quality antibodies to perform successful chromatin immunoprecipitation (ChIP) experiments. When investigating the protein-DNA interaction of interest, valid and reliable results simply cannot be attained using nonspecific and inefficient antibodies. If the antibody doesn’t come “ChIP-qualified” or “ChIP-grade” from the supplier, there are several tips you can follow to determine if it’s likely to perform well in ChIP and won’t pull down distracting material to get in the way of a successful study. Carry out the standard [more…]