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


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

STAR Protocols, cilt.3, sa.4, 2022 (Scopus) identifier identifier

  • 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: Scopus
  • Anahtar Kelimeler: Cell Biology, Cell Isolation, Mass Spectrometry, Protein Biochemistry, Proteomics
  • Abdullah Gül Üniversitesi Adresli: Evet

Özet

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.