Chromatrap has launched a protocol designed 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.
Native ChIP assays have traditionally involved five key time-consuming steps: preparation of chromatin to be analysed 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 N-ChIP kit uses discs of an inert, porous, polymer to which Protein A or Protein G has been covalently attached to maximise the capture efficiency of the target chromatin/antibody complex.
It eliminates the use of agarose or magnetic beads, which means that more sample is retained during the extraction process.
In addition, the affinity of antibody binding to antigens on Native chromatin is increased as peptides are more accessible.
