Immunostaining with modified histone antibodies

Chromatin undergoes remodelling in a variety of ways, to activate or restrict gene transcription. Histone modification is central to this. Modified histone antibodies are widely used in immunostaining and immunolabeling assays, to study the complex pathways involved. Recently, E. Terrenoire et al used a new antibody immunostaining method to define features of the human epigenome at the single-cell level.

The histone antibodies purchased from antibody suppliers are used for a wide range of assay techniques. In recent years new and improved technologies have opened exciting new avenues of research. One of these is Chromatin Immunoprecipitation, or ChIP, which examines the interaction between DNA and specific cellular proteins, working on the principle that DNA-binding proteins can be chemically cross-linked to the DNA itself. By using specific antibodies, the protein-DNA complex can be precipitated out of the lysate.

ChIP assays were first used in the 1980s, but there have been a number of new developments in recent years. These include ChIP-on-ChIP, Matrix ChIP and ChIP sequencing. In 2010, Terrenoire and her team used the latest ChIP technology to identify and map the individual components of human chromosomes in single human lymphoblastoid cells and constructed karyotypes undergoing metaphase. Although ChIP antibody assays had previously been used to probe the genomic action of modified histones, there had been no real attempt to explore the potential of ChIP sequencing for epigenome mapping in living cells.

With antibody suppliers providing the reagents, the scientists used a variety of immunofluorescence, immunostaining, microscopy and sequencing methods to define the distribution of histone modifications in both normal whole cells and derived chromosome sets (karyotypes).

By using antibodies to observe cells in metaphase,specific histone modifications were seen to work in combination along the euchromatic chromosome arms,with extensive remodelling of the epigenome taking place. Certain histone modifications were distributed at high levels during interphase,suggesting modified histones may influence inheritance of some genetic modifications.We at Novus Biologicals are one of the leading antibody suppliers for genetic research.

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