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
Descripción
Sumario: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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