We need to stimulate monocytes by LPS (10 ng) with brefeldin A and monoensin for 4 hrs ( TNF-alpha expression) and 24hrs (IL-10 expression). we are use the whole blood protocol.
Hi Julio, TNFa will be detectable within the supernatant within 6 hours of stimulation, IL-10 will take a minimum of 18 hours. If using Brefeldin A to keep everything internal for flow purposes I would suggest a time course of 6/12/24 hours to test. Be careful with your permeabilisation method and good luck!