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- Title
Study on Tissue Homogenization Buffer Composition for Brain Mass Spectrometry-Based Proteomics.
- Authors
Karpiński, Adam Aleksander; Torres Elguera, Julio Cesar; Sanner, Anne; Konopka, Witold; Kaczmarek, Leszek; Winter, Dominic; Konopka, Anna; Bulska, Ewa
- Abstract
Mass spectrometry-based proteomics aims to study the proteome both qualitatively and quantitatively. A key step in proteomic analysis is sample preparation, which is crucial for reliable results. We investigated the effect of the composition of the homogenization buffer used to extract proteins from brain tissue on the yield of protein extraction and the number and type of extracted proteins. Three different types of buffers were compared—detergent-based buffer (DB), chaotropic agent-based buffer (CAB) and buffer without detergent and chaotropic agent (DFB). Based on label-free quantitative protein analysis, detergent buffer was identified as the most suitable for global proteomic profiling of brain tissue. It allows the most efficient extraction of membrane proteins, synaptic and synaptic membrane proteins along with ribosomal, mitochondrial and myelin sheath proteins, which are of particular interest in the field of neurodegenerative disorders research.
- Subjects
PROTEOMICS; MEMBRANE proteins; MYELIN proteins; MYELIN sheath; RIBOSOMAL proteins
- Publication
Biomedicines, 2022, Vol 10, Issue 10, pN.PAG
- ISSN
2227-9059
- Publication type
Article
- DOI
10.3390/biomedicines10102466