6.4. Peptide Pulsing

This is a suggested peptide pulsing protocol. For more accurate instructions, please refer to your peptide manufacturers’ guidelines.

1. Collect cells and wash twice with serum-free RPMI1640 medium (SF-RPMI). Resuspend cells in SF-RPMI at 1 × 10⁶ cells/ml.
2. Add an appropriate amount of peptide (serial dilution may be used to find the best concentration) and incubate at 37°C, 5% CO₂ for 90 min.
3. Wash once with fresh complete RPMI medium before resuspending at the appropriate concentration for coculture (See Product Certificate for Cat.# YJUR-TCMV-PBle).