Work

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

Year: 2021

Type: article

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) 9. Mair, F. ∙ Erickson, J.R. ∙ Voillet, V. ... A targeted multi-omic analysis approach measures protein expression and low-abundance transcripts on the single-cell level Cell Rep. 2020; 31 :107499 Google Scholar .

Source: STAR Protocols

DOI: 10.1016/j.xpro.2021.100893

Authors: Xiaoshan Shi, Gisele V. Baracho, Woodrow E. Lomas, Stephanie J. Widmann, Aaron J. Tyznik

Institution: BD Biosciences (United States)

Language: English

Topic: Single-cell and spatial transcriptomics

Subfield: Molecular Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

SDG: Good health and well-being

Open Access status: gold

Estimated APC paid: $1,500