We use cookies to provide visitors of our website with the best possible experience. To learn more how we use cookies or how to block cookies, please visit our cookie policy.

Resources - Scientific Posters

BacMam-Enabled Cellular Assays Measuring Histone Posttranslational Modifications (Presentation)


May 05, 2009


Authors: Roland Leathers, Kevin Lowitz, Coby Carlson, Thomas Machleidt, Kun Bi, Justin Wetter and Matthew Robers, Invitrogen, Madison, Wisconsin; Peter Banks, BioTek Instruments, Winooski, VT



The eukaryotic nucleosome, composed of histones H2A, H2B, H3, and H4, regulates the structure of chromatin and consequently modulates gene transcription profiles in a concerted manner. Nucleosome function is directly regulated by a multitude of posttranslational modifications on amino-terminal tails of core histones, including acetylation, phosphorylation, methylation and ubiquitination. These modifications occur in a tightly regulated fashion, and affect numerous cellular processes including histone deposition, chromatin assembly, and chromosome condensation during both mitosis and meiosis. Therefore, a number of histone modifying enzymes have been identified as valuable targets for therapeutic intervention. The combination of Baculovirusmediated gene delivery (BacMam) with LanthaScreen® cellular assay technology and measurements using the BioTek Synergy™ 4 Hybrid Multi-Mode Microplate Reader, enables a powerful platform for the analysis of target-specific posttranslational modifications of histones in the cell line of interest. Specifically, we have developed HTS-compatible cellular assays measuring acetylation of histone H3 at Lys9, phosphorylation of histone H3 at Ser10, and the ubiquitination status of histone H2B. These assays together with sensitive filter-based detection on the Synergy™ 4 represent a flexible method to measure the ability of compounds to affect histone posttranslational modifications in a HTS-compatible format.