Enrichment of antigen-specific CD8+ T cell precursors or LCMV-specific CD8+ T cells at memory time points was performed as previously described (Haluszczak et al., 2009 (link)). Combined spleen and lymph nodes or spleen only, as indicated, was digested with collagenase D. Cells were labeled with PE- or APC-conjugated tetramers, followed by magnetic enrichment over columns using anti-PE or anti-APC magnetic beads (Miltenyi Biotec). Enriched samples were stained with surface antibodies, and AccuCheck counting beads (Invitrogen) were used for calculating cell number.
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Enrichment of Antigen-Specific CD8+ T Cells
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Cd8 antigen
Cells
Collagenase
Lcmv
Lymph nodes
Memory t cells
Spleen
Surface antibodies
T cell precursors
Tetramers
Corresponding Organization : University of Minnesota Medical Center
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Variable analysis
independent variables
- Enrichment of antigen-specific CD8+ T cell precursors or LCMV-specific CD8+ T cells at memory time points
- Cell number of enriched samples
- Combined spleen and lymph nodes or spleen only, as indicated
- Collagenase D digestion
- PE- or APC-conjugated tetramers labeling
- Magnetic enrichment over columns using anti-PE or anti-APC magnetic beads
- Surface antibodies staining
- AccuCheck counting beads (Invitrogen) for calculating cell number
- Positive control: Not explicitly mentioned
- Negative control: Not explicitly mentioned
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