Co-staining human PBMCs with fluorescent antibodies and antibody-oligonucleotide conjugates for cell sorting prior to single-cell CITE-Seq

Xiaoshan Shi; Gisele V Baracho; Woodrow E Lomas 3rd; Stephanie J Widmann; Aaron J Tyznik

doi:10.1016/j.xpro.2021.100893

Co-staining human PBMCs with fluorescent antibodies and antibody-oligonucleotide conjugates for cell sorting prior to single-cell CITE-Seq

STAR Protoc. 2021 Oct 19;2(4):100893. doi: 10.1016/j.xpro.2021.100893. eCollection 2021 Dec 17.

Authors

Xiaoshan Shi¹, Gisele V Baracho², Woodrow E Lomas 3rd¹, Stephanie J Widmann², Aaron J Tyznik²

Affiliations

¹ BD Biosciences, 2350 Qume Drive, San Jose, CA 95131, USA.
² BD Biosciences, 11077 N Torrey Pines Road, La Jolla, CA 92037, USA.

Abstract

The dual interrogation of the transcriptome and proteome with single-cell resolution provides exquisite insights into immune mechanisms in health and disease. Here, we describe a cutting-edge protocol wherein we combine Cellular Indexing of Transcriptomes and Epitopes by Sequencing (CITE-Seq), a technique utilizing antibody-oligonucleotide conjugates (AOCs), with fluorescence-activated cell sorting to enrich rare cell populations. Our protocol incorporates co-staining of cells with both fluorescent antibodies and AOCs simultaneously for subsequent input into the cell sorting and CITE-Seq pipeline. For complete details on the use and execution of this protocol, please refer to Mair et al. (2020).

Keywords: Antibody; Biotechnology and bioengineering; Cell isolation; Flow Cytometry/Mass Cytometry; Immunology; Molecular Biology; Single Cell.

MeSH terms

Epitopes / chemistry
Flow Cytometry / methods*
Fluorescent Antibody Technique / methods*
Gene Expression Profiling / methods*
Humans
Leukocytes, Mononuclear* / chemistry
Leukocytes, Mononuclear* / cytology
Single-Cell Analysis / methods*

Substances

Epitopes