Co-staining human PBMCs with fluorescent antibodies and antibody-oligonucleotide conjugates for cell sorting prior to single-cell CITE-Seq
Summary: 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-Se...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2021-12-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166721005992 |
| _version_ | 1818873810825248768 |
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| author | Xiaoshan Shi Gisele V. Baracho Woodrow E. Lomas, III Stephanie J. Widmann Aaron J. Tyznik |
| author_facet | Xiaoshan Shi Gisele V. Baracho Woodrow E. Lomas, III Stephanie J. Widmann Aaron J. Tyznik |
| author_sort | Xiaoshan Shi |
| collection | DOAJ |
| description | Summary: 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). |
| first_indexed | 2024-12-19T13:00:38Z |
| format | Article |
| id | doaj.art-a0c6a615e0444b528e8556f46de70088 |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-12-19T13:00:38Z |
| publishDate | 2021-12-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-a0c6a615e0444b528e8556f46de700882022-12-21T20:20:15ZengElsevierSTAR Protocols2666-16672021-12-0124100893Co-staining human PBMCs with fluorescent antibodies and antibody-oligonucleotide conjugates for cell sorting prior to single-cell CITE-SeqXiaoshan Shi0Gisele V. Baracho1Woodrow E. Lomas, III2Stephanie J. Widmann3Aaron J. Tyznik4BD Biosciences, 2350 Qume Drive, San Jose, CA 95131, USABD Biosciences, 11077 N Torrey Pines Road, La Jolla, CA 92037, USABD Biosciences, 2350 Qume Drive, San Jose, CA 95131, USABD Biosciences, 11077 N Torrey Pines Road, La Jolla, CA 92037, USABD Biosciences, 11077 N Torrey Pines Road, La Jolla, CA 92037, USA; Corresponding authorSummary: 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).http://www.sciencedirect.com/science/article/pii/S2666166721005992Cell isolationSingle CellFlow Cytometry/Mass CytometryImmunologyMolecular BiologyAntibody |
| spellingShingle | Xiaoshan Shi Gisele V. Baracho Woodrow E. Lomas, III Stephanie J. Widmann Aaron J. Tyznik Co-staining human PBMCs with fluorescent antibodies and antibody-oligonucleotide conjugates for cell sorting prior to single-cell CITE-Seq STAR Protocols Cell isolation Single Cell Flow Cytometry/Mass Cytometry Immunology Molecular Biology Antibody |
| title | Co-staining human PBMCs with fluorescent antibodies and antibody-oligonucleotide conjugates for cell sorting prior to single-cell CITE-Seq |
| title_full | Co-staining human PBMCs with fluorescent antibodies and antibody-oligonucleotide conjugates for cell sorting prior to single-cell CITE-Seq |
| title_fullStr | Co-staining human PBMCs with fluorescent antibodies and antibody-oligonucleotide conjugates for cell sorting prior to single-cell CITE-Seq |
| title_full_unstemmed | Co-staining human PBMCs with fluorescent antibodies and antibody-oligonucleotide conjugates for cell sorting prior to single-cell CITE-Seq |
| title_short | Co-staining human PBMCs with fluorescent antibodies and antibody-oligonucleotide conjugates for cell sorting prior to single-cell CITE-Seq |
| title_sort | co staining human pbmcs with fluorescent antibodies and antibody oligonucleotide conjugates for cell sorting prior to single cell cite seq |
| topic | Cell isolation Single Cell Flow Cytometry/Mass Cytometry Immunology Molecular Biology Antibody |
| url | http://www.sciencedirect.com/science/article/pii/S2666166721005992 |
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