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Purification and quantitative proteomic analysis of cell bodies and protrusions
Actin-rich protrusions are membrane extensions generated by actin polymerization that drive mesenchymal-like cell migration. Characterization of protrusions proteome is crucial for understanding their function. We present a complete step-by-step protocol based on microporous filter-based fractionati...
Autores principales: | , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8065254/ https://www.ncbi.nlm.nih.gov/pubmed/33912849 http://dx.doi.org/10.1016/j.xpro.2021.100462 |
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author | Dermit, Maria Mardakheh, Faraz K. |
author_facet | Dermit, Maria Mardakheh, Faraz K. |
author_sort | Dermit, Maria |
collection | PubMed |
description | Actin-rich protrusions are membrane extensions generated by actin polymerization that drive mesenchymal-like cell migration. Characterization of protrusions proteome is crucial for understanding their function. We present a complete step-by-step protocol based on microporous filter-based fractionation of protrusive cellular domains coupled with sample preparation for quantitative proteomics, mass spectrometric data acquisition, and data analysis. This protocol enables purification, quantification, and analysis of the distribution of proteins present in protrusions and cell bodies. For complete details on the use and execution of this protocol, please refer to Dermit et al. (2020). |
format | Online Article Text |
id | pubmed-8065254 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
spelling | pubmed-80652542021-04-27 Purification and quantitative proteomic analysis of cell bodies and protrusions Dermit, Maria Mardakheh, Faraz K. STAR Protoc Protocol Actin-rich protrusions are membrane extensions generated by actin polymerization that drive mesenchymal-like cell migration. Characterization of protrusions proteome is crucial for understanding their function. We present a complete step-by-step protocol based on microporous filter-based fractionation of protrusive cellular domains coupled with sample preparation for quantitative proteomics, mass spectrometric data acquisition, and data analysis. This protocol enables purification, quantification, and analysis of the distribution of proteins present in protrusions and cell bodies. For complete details on the use and execution of this protocol, please refer to Dermit et al. (2020). Elsevier 2021-04-10 /pmc/articles/PMC8065254/ /pubmed/33912849 http://dx.doi.org/10.1016/j.xpro.2021.100462 Text en © 2021 The Authors https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Protocol Dermit, Maria Mardakheh, Faraz K. Purification and quantitative proteomic analysis of cell bodies and protrusions |
title | Purification and quantitative proteomic analysis of cell bodies and protrusions |
title_full | Purification and quantitative proteomic analysis of cell bodies and protrusions |
title_fullStr | Purification and quantitative proteomic analysis of cell bodies and protrusions |
title_full_unstemmed | Purification and quantitative proteomic analysis of cell bodies and protrusions |
title_short | Purification and quantitative proteomic analysis of cell bodies and protrusions |
title_sort | purification and quantitative proteomic analysis of cell bodies and protrusions |
topic | Protocol |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8065254/ https://www.ncbi.nlm.nih.gov/pubmed/33912849 http://dx.doi.org/10.1016/j.xpro.2021.100462 |
work_keys_str_mv | AT dermitmaria purificationandquantitativeproteomicanalysisofcellbodiesandprotrusions AT mardakhehfarazk purificationandquantitativeproteomicanalysisofcellbodiesandprotrusions |