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Tandem affinity purification protocol for isolation of protein complexes from Schizosaccharomyces pombe

Many cellular processes require the activities of complex molecular machines composed of several protein subunits. Insights into these systems can be gained by isolation of protein complexes followed by in vitro analyses determining the identity, posttranslational modifications, and interactions amo...

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Autores principales: Cipak, Lubos, Selicky, Tomas, Jurcik, Jan, Cipakova, Ingrid, Osadska, Michaela, Lukacova, Veronika, Barath, Peter, Gregan, Juraj
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8808283/
https://www.ncbi.nlm.nih.gov/pubmed/35128479
http://dx.doi.org/10.1016/j.xpro.2022.101137
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author Cipak, Lubos
Selicky, Tomas
Jurcik, Jan
Cipakova, Ingrid
Osadska, Michaela
Lukacova, Veronika
Barath, Peter
Gregan, Juraj
author_facet Cipak, Lubos
Selicky, Tomas
Jurcik, Jan
Cipakova, Ingrid
Osadska, Michaela
Lukacova, Veronika
Barath, Peter
Gregan, Juraj
author_sort Cipak, Lubos
collection PubMed
description Many cellular processes require the activities of complex molecular machines composed of several protein subunits. Insights into these systems can be gained by isolation of protein complexes followed by in vitro analyses determining the identity, posttranslational modifications, and interactions among proteins. Here, we present a protocol for tandem affinity purification (TAP) of protein complexes from the fission yeast Schizosaccharomyces pombe. The protocol employs cells expressing C-terminally TAP-tagged proteins and is suitable for the analysis of purified proteins by mass spectrometry. For complete information on the use and execution of this protocol, please refer to Cipakova et al. (2019).
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spelling pubmed-88082832022-02-04 Tandem affinity purification protocol for isolation of protein complexes from Schizosaccharomyces pombe Cipak, Lubos Selicky, Tomas Jurcik, Jan Cipakova, Ingrid Osadska, Michaela Lukacova, Veronika Barath, Peter Gregan, Juraj STAR Protoc Protocol Many cellular processes require the activities of complex molecular machines composed of several protein subunits. Insights into these systems can be gained by isolation of protein complexes followed by in vitro analyses determining the identity, posttranslational modifications, and interactions among proteins. Here, we present a protocol for tandem affinity purification (TAP) of protein complexes from the fission yeast Schizosaccharomyces pombe. The protocol employs cells expressing C-terminally TAP-tagged proteins and is suitable for the analysis of purified proteins by mass spectrometry. For complete information on the use and execution of this protocol, please refer to Cipakova et al. (2019). Elsevier 2022-01-28 /pmc/articles/PMC8808283/ /pubmed/35128479 http://dx.doi.org/10.1016/j.xpro.2022.101137 Text en © 2022 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
Cipak, Lubos
Selicky, Tomas
Jurcik, Jan
Cipakova, Ingrid
Osadska, Michaela
Lukacova, Veronika
Barath, Peter
Gregan, Juraj
Tandem affinity purification protocol for isolation of protein complexes from Schizosaccharomyces pombe
title Tandem affinity purification protocol for isolation of protein complexes from Schizosaccharomyces pombe
title_full Tandem affinity purification protocol for isolation of protein complexes from Schizosaccharomyces pombe
title_fullStr Tandem affinity purification protocol for isolation of protein complexes from Schizosaccharomyces pombe
title_full_unstemmed Tandem affinity purification protocol for isolation of protein complexes from Schizosaccharomyces pombe
title_short Tandem affinity purification protocol for isolation of protein complexes from Schizosaccharomyces pombe
title_sort tandem affinity purification protocol for isolation of protein complexes from schizosaccharomyces pombe
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8808283/
https://www.ncbi.nlm.nih.gov/pubmed/35128479
http://dx.doi.org/10.1016/j.xpro.2022.101137
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