Question: How can I combine dextramer staining and cell surface protein staining in the same workflow?
Answer: When combining dextramer staining and cell surface protein staining protocols, the most important workflow consideration is to make sure that the dextramer staining is performed prior to antibody staining. The cells should be stained with the dextramer pool for 10 minutes at 4C without light exposure; the antibody pool can then be added afterward (at this step, the cells should be stained for 30 minutes at 4C).
Please see our protocol for Cell Labeling with Dextramers, which also includes modifications necessary for incorporating cell surface protein staining, for more detailed instructions.
This step crucial to obtaining high-quality dextramer data, since dextramer interactions with the TCRs are not as strong as antibody-epitope interactions, and thus may be overshadowed if the staining is performed simultaneously.
Post-staining, the 10x workflow should be executed as normal according to our Single Cell V(D)J Reagent Kits User Guide with Feature Barcoding technology for Cell Surface Protein. There are no other modifications necessary to the workflow. Dextramer and Cell Surface Protein oligos will be incorporated into the same final library during the workflow, and Cell Ranger will separate them based on the barcodes detected.
Products: Single Cell Immune Profiling