Protocol for cell surface biotinylation of magnetic labeled and captured human peripheral blood mononuclear cells

Analysis of the surfaceome of a blood cell subset requires cell sorting, followed by surface protein enrichment. Here, we present a protocol combining magnetically activated cell sorting (MACS) and surface biotinylation of the target cell subset from human peripheral blood mononuclear cells (PBMCs)....

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Detalles Bibliográficos
Autores principales: Ayaz-Guner, Serife, Acar, Mustafa Burak, Boyvat, Dudu, Guner, Huseyin, Bozalan, Habibe, Güzel, Melis, Yıldır, Selin Kübra, Altınsoy, Nilay, Fındık, Fatma, Karakükçü, Musa, Özcan, Servet
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9692030/
https://www.ncbi.nlm.nih.gov/pubmed/36595912
http://dx.doi.org/10.1016/j.xpro.2022.101863
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author Ayaz-Guner, Serife
Acar, Mustafa Burak
Boyvat, Dudu
Guner, Huseyin
Bozalan, Habibe
Güzel, Melis
Yıldır, Selin Kübra
Altınsoy, Nilay
Fındık, Fatma
Karakükçü, Musa
Özcan, Servet
author_facet Ayaz-Guner, Serife
Acar, Mustafa Burak
Boyvat, Dudu
Guner, Huseyin
Bozalan, Habibe
Güzel, Melis
Yıldır, Selin Kübra
Altınsoy, Nilay
Fındık, Fatma
Karakükçü, Musa
Özcan, Servet
author_sort Ayaz-Guner, Serife
collection PubMed
description Analysis of the surfaceome of a blood cell subset requires cell sorting, followed by surface protein enrichment. Here, we present a protocol combining magnetically activated cell sorting (MACS) and surface biotinylation of the target cell subset from human peripheral blood mononuclear cells (PBMCs). We describe the steps for isolating target cells and their in-column surface biotinylation, followed by isolation and mass spectrometry analysis of biotinylated proteins. The protocol enables in-column surface biotinylation of specific cell subsets with minimal membrane disruption.
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spelling pubmed-96920302022-11-26 Protocol for cell surface biotinylation of magnetic labeled and captured human peripheral blood mononuclear cells Ayaz-Guner, Serife Acar, Mustafa Burak Boyvat, Dudu Guner, Huseyin Bozalan, Habibe Güzel, Melis Yıldır, Selin Kübra Altınsoy, Nilay Fındık, Fatma Karakükçü, Musa Özcan, Servet STAR Protoc Protocol Analysis of the surfaceome of a blood cell subset requires cell sorting, followed by surface protein enrichment. Here, we present a protocol combining magnetically activated cell sorting (MACS) and surface biotinylation of the target cell subset from human peripheral blood mononuclear cells (PBMCs). We describe the steps for isolating target cells and their in-column surface biotinylation, followed by isolation and mass spectrometry analysis of biotinylated proteins. The protocol enables in-column surface biotinylation of specific cell subsets with minimal membrane disruption. Elsevier 2022-11-21 /pmc/articles/PMC9692030/ /pubmed/36595912 http://dx.doi.org/10.1016/j.xpro.2022.101863 Text en © 2022 The Authors https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Ayaz-Guner, Serife
Acar, Mustafa Burak
Boyvat, Dudu
Guner, Huseyin
Bozalan, Habibe
Güzel, Melis
Yıldır, Selin Kübra
Altınsoy, Nilay
Fındık, Fatma
Karakükçü, Musa
Özcan, Servet
Protocol for cell surface biotinylation of magnetic labeled and captured human peripheral blood mononuclear cells
title Protocol for cell surface biotinylation of magnetic labeled and captured human peripheral blood mononuclear cells
title_full Protocol for cell surface biotinylation of magnetic labeled and captured human peripheral blood mononuclear cells
title_fullStr Protocol for cell surface biotinylation of magnetic labeled and captured human peripheral blood mononuclear cells
title_full_unstemmed Protocol for cell surface biotinylation of magnetic labeled and captured human peripheral blood mononuclear cells
title_short Protocol for cell surface biotinylation of magnetic labeled and captured human peripheral blood mononuclear cells
title_sort protocol for cell surface biotinylation of magnetic labeled and captured human peripheral blood mononuclear cells
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9692030/
https://www.ncbi.nlm.nih.gov/pubmed/36595912
http://dx.doi.org/10.1016/j.xpro.2022.101863
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