This protocol describes the comparative proteomic profiling of the spleen of wild type versus 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).
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http://dx.doi.org/10.1016/j.xpro.2020.100196 | DOI Listing |
J Proteome Res
January 2025
Department of Medicine and Surgery, Proteomics and Metabolomics Unit, University of Milano-Bicocca, Vedano al Lambro 20854, Italy.
MALDI-HiPLEX-IHC mass spectrometry imaging (MSI) represents a newly established workflow to map tens of antibodies linked to photocleavable mass tags (PC-MTs), which report the distribution of antigens in formalin-fixed paraffin-embedded (FFPE) tissue sections. While this highly multiplexed approach has previously been integrated with untargeted methods, the possibility of mapping target cell antigens and performing bottom-up spatial proteomics on the same tissue section has yet to be explored. This proof-of-concept study presents a novel workflow combining MALDI-HiPLEX-IHC with untargeted spatial proteomics to analyze a single FFPE tissue section, using clinical clear cell renal cell carcinoma (ccRCC) tissue as a model.
View Article and Find Full Text PDFAlzheimers Dement
December 2024
Florey Institute of Neuroscience and Mental Health, University of Melbourne, Parkville, VIC, Australia.
Background: A better understanding of the molecular process that drive Alzheimer's disease(AD) are required to develop effective biomarkers and therapies. This includes determining how essential elements like Fe, Cu and Zn are involved in the disease. In the literature there is debate over the role of iron in AD and there are reports of increased, decreased and unchanged levels of Fe in AD brain.
View Article and Find Full Text PDFTrans R Soc Trop Med Hyg
January 2025
Instituto Clodomiro Picado, Facultad de Microbiología, Universidad de Costa Rica, San José 11501, Costa Rica.
Background: The genus Metlapilcoatlus was recently erected to include six species of stout venomous snakes, known as the jumping pitvipers, which inhabit mountainous areas of Mesoamerica. This group maintains affinity with Atropoides picadoi, another jumping pitviper with restricted distribution in Costa Rica and Panama. Although the venom of A.
View Article and Find Full Text PDFMethods Mol Biol
December 2024
Department of Chemistry, North Carolina State University, Raleigh, NC, USA.
Extracellular matrix (ECM) from decellularized mammalian tissues has been used in many therapeutic applications. The tissue-specific composition of the ECM is critically associated with therapeutic performance. However, ECM translation needs to be improved because of the complex composition and limited understanding of ECM repairing mechanisms due partly to incomplete proteomic interrogation of ECM samples.
View Article and Find Full Text PDFMethods Mol Biol
December 2024
Department of Chemistry, North Carolina State University, Raleigh, NC, USA.
In recent years, discovery proteomics has emerged as a pivotal tool in biological research, especially when studying the intricate relationships among multiple organisms. To delve deeper into these interactions, we pioneered a bottom-up proteomics workflow. Using nanoLC-MS/MS and a label-free quantification method, this work specifically examines the differential protein expression in fleas (Ctenocephalides felis felis) that have been experimentally infected with Bartonella henselae, the causative agent of cat scratch disease (CSD).
View Article and Find Full Text PDFEnter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!