Tetramer enrichment was used to isolate antigen-specific cells from pre-immune or acutely challenged mice. A modification of the method used by Obar et al., 2008 (link) was employed. Following digestion with collagenase D, single-cell suspensions were prepared from the spleens (acutely challenged mice) or spleen and cervical, axillary, brachial, inguinal, and mesenteric lymph nodes (pre-immune mice). When possible, the same tetramer (Trp2180–188/Kb) was used in both APC and PE to ensure specificity. Anti-PE and anti-APC beads and magnetized columns (both from Miltenyi Biotec) were used to enrich for tetramer-bound cells. Samples were stained and analyzed by flow cytometry; CountBright counting beads (Invitrogen) were used for enumeration.
Anti pe and anti apc beads
Manufactured by Miltenyi Biotec
Anti-PE and anti-APC beads are magnetic beads coated with antibodies that specifically bind to phycoerythrin (PE) or allophycocyanin (APC) fluorescent dyes. These beads can be used to separate and enrich cells or molecules labeled with PE or APC, facilitating downstream applications such as flow cytometry or cell sorting.
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Lab products found in correlation
2 protocols using anti pe and anti apc beads
1
Tetramer-based Enrichment of Antigen-specific Cells
2
Antigen-specific T cell enrichment
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Tetramer enrichment was used to isolate antigen-specific cells from pre-immune or acutely challenged mice. A modification of the method used by Obar et al. (Obar et al., 2008) (link) was employed. Following digestion with collagenase D, single-cell suspensions were prepared from the spleens (acutely challenged mice) or spleen and peripheral/mesenteric lymph nodes (pre-immune mice). When possible, the same tetramer (Trp2180-188/K b ) was used in both APC and PE to ensure specificity. Anti-PE and anti-APC beads and magnetized columns (both from Miltenyi Biotec) were used to enrich for tetramer-bound cells. Samples were stained and analyzed by flow cytometry; CountBright counting beads (Invitrogen) were used for enumeration.
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