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., ...More

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

  • 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
  • Kayseri University Affiliated: No

Abstract

© 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.