HDAC/HAT Function Assessment and Inhibitor Development

0578923 - BTÚ 2024 RIV US eng M - Část monografie knihy
Mikešová, Jana - Ondráková, Markéta - Jelínková, Iva - Ptáček, Jakub - Nováková, Zora - Bařinka, Cyril
Determining Potency of Inhibitors Targeting Histone Deacetylase 6 by Quantification of Acetylated Tubulin in Cells.
HDAC/HAT Function Assessment and Inhibitor Development. Vol. 2589. New York: Springer Nature, 2023, s. 455-466. ISBN 978-1-0716-2788-4
Grant CEP: GA ČR(CZ) GA21-31806S
Institucionální podpora: RVO:86652036
Klíčová slova: Acetyl-histone * Acetyl-tubulin * Monoclonal antibody 6-11B-1 * Quantitative Western blotting
Obor OECD: Biochemistry and molecular biology
https://link.springer.com/protocol/10.1007/978-1-0716-2788-4_29

During the preclinical development of small molecule inhibitors, compounds or compound libraries are typically first screened using purified target enzymes in vitro to select candidates with high potency. In the later stages of the development, however, functional cell-based assays may provide biologically more relevant data. In this chapter, we describe a detailed protocol for determining the potency of inhibitors targeting human histone deacetylase 6 in complex cellular environments. Cells are first treated with a dilution series of tested compounds, cell lysates separated by SDS-PAGE, and electrotransferred to a blotting membrane. The inhibitor potency is then determined indirectly by quantifying the levels of acetylated tubulin as a surrogate readout.
Trvalý link: https://hdl.handle.net/11104/0347838