Hi Mónica Itzel Martínez GutiérrezYes, that approach works. You can dilute the buffy coat 1:1 with PBS or HBSS, then carefully layer about 35 mL of the diluted sample over 15 mL of Ficoll in a 50 mL tube. From there, follow the standard PBMC isolation protocol.

A few key reminders:

• Ensure both the Ficoll and your PBS/HBSS are at room temperature.
• Handle the layering step gently to avoid mixing.
• Centrifuge according to your usual PBMC protocol to obtain a clean interface.

For a more detailed explanation and troubleshooting tips, including PBMC isolation from buffy coats, the HANC Cross-Network PBMC Processing SOP is an excellent reference.

Good Luck!