Protocol for the Bottom-Up Proteomic Analysis of Mouse Spleen.

1. STAR Protoc. 2020 Dec 3;1(3):100196. doi: 10.1016/j.xpro.2020.100196. eCollection 2020 Dec 18. Protocol for the Bottom-Up Proteomic Analysis of Mouse Spleen. Dowling P(1)(2), Gargan S(1)(2), Zweyer M(3), Henry M(4), Meleady P(4), Swandulla D(5), Ohlendieck K(1)(2). Author information: (1)Department of Biology, Maynooth University, National University of Ireland, Maynooth W23F2H6, Co. Kildare, Ireland. (2)Kathleen Lonsdale Institute for Human Health Research, Maynooth University, Maynooth W23F2H6, Co. Kildare, Ireland. (3)Department of Neonatology and Paediatric Intensive Care, Children's Hospital, University of Bonn, D53113 Bonn, Germany. (4)National Institute for Cellular Biotechnology, Dublin City University, Dublin 9, Ireland. (5)Institute of Physiology II, University of Bonn, D53115 Bonn, Germany. This protocol describes the comparative proteomic profiling of the spleen of wild type versus mdx-4cv mouse, a model of dystrophinopathy. We detail sample preparation for bottom-up proteomic mass spectrometry experiments, including homogenization of tissue, protein concentration measurements, protein digestion, and removal of interfering chemicals. We then describe the steps for mass spectrometric analysis and bioinformatic evaluation. For complete details on the use and execution of this protocol, please refer to Dowling et al. (2020). © 2020 The Author(s). DOI: 10.1016/j.xpro.2020.100196 PMCID: PMC7757555 PMID: 33377090 [Indexed for MEDLINE] Conflict of interest statement: The authors declare no competing interests.