ABSTRACT
Epigenetic programming plays a vital role in regulating pluripotency genes, which become activated or inactivated during the processes of dedifferentiation and differentiation during an organism's development. The analysis of epigenetic modifications has become possible through the technique of immunostaining, where specific antibodies allow the identification of a single target protein. This chapter describes a detailed protocol for the analysis of the epigenetic modifications with the use of confocal microscopy, subsequent image, and statistical analysis on the example of Fagopyrum calli with the use of nine antibodies raised against histone H3 and H4 methylation and acetylation on several lysines as well as DNA methylation.
Subject(s)
Fagopyrum , Fagopyrum/metabolism , Histones/metabolism , Cell Nucleus/metabolism , DNA Methylation , Antibodies/metabolism , Epigenesis, Genetic , AcetylationABSTRACT
Immunostaining is a well-established technique for identifying specific proteins in tissue samples with specific antibodies to identify a single target protein. It is commonly used in research and provides information about cellular localization and protein expression levels. This chapter describes a detailed protocol for immunostaining fixed Fagopyrum calli embedded in Steedman's wax using nine antibodies raised against histone H3 and H4 methylation and acetylation on several lysines and DNA methylation.