Simple & Efficient Protocol for Native Chromatin Immunoprecipitation

Simple & Efficient Protocol for Native Chromatin Immunoprecipitation

22 Jan 2016
Alex Waite
Editorial Assistant

Chromatrap, a business unit of Porvair Sciences, announces a new patented protocol that is able to simply, quickly and efficiently enrich transcription factors from Native Chromatin.

Native Chromatin Immunoprecipitation (N-ChIP) is an alternative to traditional cross-linked (X-ChIP), which removes the need for fixing cells with formaldehyde prior to extraction. As a consequence, the cells remain in a more natural 'native state'. The DNA/protein fragments are selectively immunoprecipitated using antibodies directed against the protein of interest and the resulting fractions treated to separate the DNA and protein components. Native ChIP is the technique of choice for the study of histone modifiers and certain abundant transcription factors that are likely to be bound to DNA.

Traditionally Native ChIP assays have involved five key time-consuming

steps: Preparation of chromatin to be analyzed from cells; Immunoprecipitation of chromatin using a high quality ChIP-validated antibody specific to the target protein; isolation of the precipitated chromatin fragments; DNA recovery from the precipitated product and finally DNA analysis.

The ChromatrapR N-ChIP kit provides a new quicker, easier and more efficient way of performing N-ChIP assays (Patent No. GB2482209). It uses discs of an inert, porous, polymer to which Protein A or Protein G has been covalently attached to maximize the capture efficiency of the target chromatin/antibody complex.

Unlike other approaches the ChromatrapR N-ChIP kit eliminates using agarose or magnetic beads resulting in more sample being retained during the extraction process. In addition, ChromatrapR N-ChIP technology increases the affinity of antibody binding to antigens on Native chromatin as peptides are more accessible. As a result, the ChromatrapR N-ChIP kit sets a new standard for enriching transcription factors from Native Chromatin.

For further information on ChromatrapR N-ChIP protocol please click here and to see the range of products from Porvair click here.

Chromatrap® FFPE ChIP Kit

Chromatrap

Epigenetic identification and targeting in clinical FFPE tissues is a crucial step for potential epigenetic therapeutics and personalized medicine therapies. FFPE is a standard method to archive and preserve medical tissue biopsy samples. These archives, kept in many hospitals world-wide, are a huge, rich and untapped sample resource for genetic research. Due to the cross-linking of proteins by formaldehyde, FFPE tissues present a particular challenge for ChIP analysis. In addition, the small size and delicate nature of the tissue, and the difficulty in extracting samples from paraffin, used to provide tissue support after preservation, can damage complex cellular complexes such as chromatin. ChIP assays from these samples have therefore proved time consuming and difficult.  Chromatrap® FFPE ChIP kit overcomes these issues and provides the following benefits: Provides greater flexibility and more IPs per sample Works across a range of FFPE samples, human and animal Available in 96-well plate or spin column format Works equally well with high and low abundant marks Provides sufficient DNA to perform library preps for ChIP-seq assays

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