Profiling histone posttranslational modifications and chromatin-modifying proteins by high-throughput reverse phase protein array

Epigenetic variation contributes significantly to a wide range of activities, from normal development to human diseases through alteration of chromatin organization and gene expression. Identifying and profiling changes of histone posttranslation modifications (PTMs) and their regulatory proteins are crucial for understanding diseases and for the discovery of epigenetic therapeutic agents. In this chapter, we review the application of reverse phase protein array (RPPA) technology, a robust antibody-based platform, for profiling histone PTMs and their regulators for laboratory and clinical research. The comprehensive epigenetic analysis by RPPA provides a highly sensitive and high-throughput method for epigenetic profiling on the protein level and is complementary to mass spectrometry analysis. RPPA technology is suitable for a wide range of applications and has the potential to discover and validate epigenetic states as therapeutic targets and biomarkers, thus helping advance precision medicine and providing therapeutic guidance.