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

Atıf İçin Kopyala

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

STAR Protocols, cilt.3, sa.4, 2022 (ESCI)

Yayın Türü: Makale / Tam Makale
Cilt numarası: 3 Sayı: 4
Basım Tarihi: 2022
Doi Numarası: 10.1016/j.xpro.2022.101863
Dergi Adı: STAR Protocols
Derginin Tarandığı İndeksler: Emerging Sources Citation Index (ESCI), Scopus
Anahtar Kelimeler: Cell Biology, Cell Isolation, Mass Spectrometry, Protein Biochemistry, Proteomics
Erciyes Üniversitesi Adresli: Evet

Özet

© 2022 The AuthorsAnalysis 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.