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Protocol to identify and sort antigen-specific B cells from Plasmodium-infected mice
Carolina Calôba1, Allen M Minns2, Scott E Lindner2
1Discipline of Microbiology and Immunology, Rosalind Franklin University of Medicine and Science, North Chicago, IL, USA; School of Graduate and Postdoctoral Studies, Rosalind Franklin University of Medicine and Science, North Chicago, IL, USA; Center for Cancer Cell Biology, Immunology and Infection, Rosalind Franklin University of Medicine and Science, North Chicago, IL, USA.
Here, we detail a protocol to identify an antigen-specific B cell population using SpyCage reagents tailored to bind MSP1-19 antigen in a rodent malaria model. We describe steps for the preparation of splenic single-cell suspension, B cell enrichment, and staining to facilitate sorting of live cells for downstream applications such as single-cell RNA, V(D)J, and assay for transposase-accessible chromatin (ATAC) sequencing. For complete details on the use and execution of this protocol, please refer to Calôba et al.1.
Here, we detail a protocol to identify an antigen-specific B cell population using SpyCage reagents tailored to bind MSP1-19 antigen in a rodent malaria model. We describe steps for the preparation of splenic single-cell suspension, B cell enrichment, and staining to facilitate sorting of live cells for downstream applications such as single-cell RNA, V(D)J, and assay for transposase-accessible chromatin (ATAC) sequencing. For complete details on the use and execution of this protocol, please refer to Calôba et al.1.
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