Single-Cell Protein Analysis - Methods and Protocols
Single-Cell Protein Analysis - Methods and Protocols
复制标题
单细胞蛋白质分析 - 方法和实验方案
DOI：
10.1007/978-1-0716-1771-7_13
复制
发表时间：
2022
期刊：
影响因子：
--
通讯作者：
Mende N
中科院分区：
文献类型：
--
作者：
Mende N
CITE-seq工作流程结合了传统的单细胞转录组学分析与使用抗人白细胞介素缀合抗体的细胞表面蛋白表达的同时分析。这种对mRNA数据的免疫表型分析的增加允许更详细地表征单细胞异质性，并且可以帮助鉴定用于转录定义的新细胞亚群的前瞻性分离的标记物。在此，我们描述了使用来自BioLegend的市售TotalSeq™抗体和来自10 x Genomics的基于液滴的单细胞RNA-seq商业平台制备人脐带血单核细胞和富含CD 34+的造血祖细胞以同时表征蛋白质和RNA的工作流程。
The CITE-seq workflow combines conventional single-cell transcriptomic analysis with simultaneous analysis of cell surface protein expression using oligonucleotide-conjugated antibodies. This addition of immunophenotyping to mRNA data allows for a more detailed characterization of single-cell heterogeneity and can help to identify markers for the prospective isolation of transcriptionally defined novel cell subsets. Here, we describe the workflow for the preparation of human cord blood mononuclear cells and CD34+-enriched hematopoietic progenitors for the simultaneous characterization of protein and RNA using the commercially available TotalSeq™ antibodies from BioLegend and the droplet-based single-cell RNA-seq commercial platform from 10x Genomics.