Question: Is bead-based pre-enrichment of B or T cells recommended for Barcode Enabled Antigen Mapping (BEAM)?
Answer: Bead-based pre-enrichment of B or T cells is not required but may be beneficial if the starting percentage of B or T cells is low. Performing a bead-based pre-enrichment step can reduce the number of cells needed as input into BEAM labeling and flow sorting. This may reduce flow sorting time. It may also reduce the volume of each BEAM Assembly required for sample labeling (the volume should be adjusted for every 1 million cells used for labeling).
If performing bead-based pre-enrichment, negative selection methods that generate “untouched” B or T cells are recommended, to avoid interference with BEAM reagents. Examples of negative selection kits include:
- Dynabeads Untouched Human CD8 T Cells Kit (Invitrogen PN-11348D)
- Dynabeads Untouched Mouse CD8 T Cells Kit (Invitrogen PN-11417D)
- EasySep™ Human B Cell Isolation Kit (Stemcell Technologies, PN-17954)
If performing bead-based enrichment upstream of BEAM-Ab, see: How can I reduce background PE signal for the BEAM-Ab Negative Control Assembly?
Products: Single Cell Immune Profiling, Barcode Enabled Antigen Mapping (BEAM)