A quick update: Joseph Mattapallil in my laboratory tested a variety of methods suggested by various researchers for the isolation of PBMC from cynomolgous monkeys. Joseph tested the following methods: (1) BioE's "Prepacyte" only protocol; (2) BioE's prepacyte followed with 100% Ficoll; (3) 55% percoll diluted with water; (4) 55% percoll diluted with PBS; (5) 85% ficoll + 15% water; (6) 85% ficoll + 15% PBS. #1 worked but still required RBC lysis for cleanup. #2, 3, and 4 did not work at all. #5 worked but PBMC were a bit close to RBC; the PBMC could still be cleanly removed. #6 worked like a charm, and is now our standard protocol. 9 mL of blood was diluted with 5 mL of PBS, and layered on 10 mL of 85:15 Ficoll:PBS mixture. Spin at 1000xg for 20 min (brakes off) at room temp. Thanks again to everyone who provided suggestions and insights. mr
This archive was generated by hypermail 2b29 : Sun Jan 05 2003 - 19:01:40 EST