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

Ayaz-Guner, Serife; Acar, Mustafa; Boyvat, Dudu; GÜNER, HÜSEYİN; Bozalan, Habibe; Güzel, Melis; Yıldır, Selin; Altınsoy, Nilay; Fındık, Fatma; KARAKÜKCÜ, MUSA; Özcan, Servet

doi:10.1016/j.xpro.2022.101863

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

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Ayaz-Guner S., Acar M. B., Boyvat D., GÜNER H., Bozalan H., Güzel M., ...More

STAR Protocols, vol.3, no.4, 2022 (Scopus)

Publication Type: Article / Article
Volume: 3 Issue: 4
Publication Date: 2022
Doi Number: 10.1016/j.xpro.2022.101863
Journal Name: STAR Protocols
Journal Indexes: Scopus
Keywords: Cell Biology, Cell Isolation, Mass Spectrometry, Protein Biochemistry, Proteomics
Abdullah Gül University Affiliated: Yes

Abstract

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.