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Study on Tissue Homogenization Buffer Composition for Brain Mass Spectrometry-Based Proteomics

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...

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Autores principales: Karpiński, Adam Aleksander, Torres Elguera, Julio Cesar, Sanner, Anne, Konopka, Witold, Kaczmarek, Leszek, Winter, Dominic, Konopka, Anna, Bulska, Ewa
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9598821/
https://www.ncbi.nlm.nih.gov/pubmed/36289728
http://dx.doi.org/10.3390/biomedicines10102466
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author Karpiński, Adam Aleksander
Torres Elguera, Julio Cesar
Sanner, Anne
Konopka, Witold
Kaczmarek, Leszek
Winter, Dominic
Konopka, Anna
Bulska, Ewa
author_facet Karpiński, Adam Aleksander
Torres Elguera, Julio Cesar
Sanner, Anne
Konopka, Witold
Kaczmarek, Leszek
Winter, Dominic
Konopka, Anna
Bulska, Ewa
author_sort Karpiński, Adam Aleksander
collection PubMed
description 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.
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spelling pubmed-95988212022-10-27 Study on Tissue Homogenization Buffer Composition for Brain Mass Spectrometry-Based Proteomics Karpiński, Adam Aleksander Torres Elguera, Julio Cesar Sanner, Anne Konopka, Witold Kaczmarek, Leszek Winter, Dominic Konopka, Anna Bulska, Ewa Biomedicines Article 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. MDPI 2022-10-02 /pmc/articles/PMC9598821/ /pubmed/36289728 http://dx.doi.org/10.3390/biomedicines10102466 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Karpiński, Adam Aleksander
Torres Elguera, Julio Cesar
Sanner, Anne
Konopka, Witold
Kaczmarek, Leszek
Winter, Dominic
Konopka, Anna
Bulska, Ewa
Study on Tissue Homogenization Buffer Composition for Brain Mass Spectrometry-Based Proteomics
title Study on Tissue Homogenization Buffer Composition for Brain Mass Spectrometry-Based Proteomics
title_full Study on Tissue Homogenization Buffer Composition for Brain Mass Spectrometry-Based Proteomics
title_fullStr Study on Tissue Homogenization Buffer Composition for Brain Mass Spectrometry-Based Proteomics
title_full_unstemmed Study on Tissue Homogenization Buffer Composition for Brain Mass Spectrometry-Based Proteomics
title_short Study on Tissue Homogenization Buffer Composition for Brain Mass Spectrometry-Based Proteomics
title_sort study on tissue homogenization buffer composition for brain mass spectrometry-based proteomics
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9598821/
https://www.ncbi.nlm.nih.gov/pubmed/36289728
http://dx.doi.org/10.3390/biomedicines10102466
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