3 articles published in JoVE
Preparation of Cell Extracts by Cryogrinding in an Automated Freezer Mill Ernest O. N. Phillips*1, Serena Giovinazzi*1, Sarah L. Menz*1, Yuna Son1, Akash Gunjan1 1Department of Biomedical Sciences, Florida State University College of Medicine We describe a reliable method for the preparation of whole cell extracts from yeast or other cells using a cryogenic freezer mill that minimizes degradation and denaturation of proteins. The cell extracts are suitable for purification of functional protein complexes, proteomic analyses, co-immunoprecipitation studies and detection of labile protein modifications.
Assays for Validating Histone Acetyltransferase Inhibitors Aaron R. Waddell1, Daiqing Liao1 1Department of Anatomy and Cell Biology, and UF Health Cancer Center, University of Florida College of Medicine Inhibitors of histone acetyltransferases (HATs, also known as lysine acetyltransferases), such as CBP/p300, are potential therapeutics for treating cancer. However, rigorous methods for validating these inhibitors are needed. Three in vitro methods for validation include HAT assays with recombinant acetyltransferases, immunoblotting for histone acetylation in cell culture, and ChIP-qPCR.
Extraction of Extracellular Vesicles from Whole Tissue Stephanie N. Hurwitz1, James M. Olcese1, David G. Meckes Jr.1 1Department of Biomedical Sciences, Florida State University College of Medicine Here, we provide a detailed protocol to isolate small extracellular vesicles (EVs) from whole tissues, including brain and tumor specimens. This method offers a reproducible technique to extract EVs from solid tissue for further downstream analyses.