fMLP for neutrophil recruitment protocol? - (Nov/08/2012 )
I am having trouble inducing neutrophil influx to the peritoneal cavity in mice using fMLP. I have tried 10mM and 20mM fMLP concentrations (200ul i.p.) and waiting for 2hr and 4hr post-injection to lavage the peritoneal cavity with 10ml HBSS and obtain cells. I am not seeing an increase in the number of cells migrating to the peritoneal cavity compared to a PBS i.p. control. If you have any ideas regarding why my protocol isn't working, or suggestions to improve it, I would greatly appreciate the help!
I don't know why the fMLP isn't working but I can tell you that if you use thioglycollate medium you can get neutrophils. 2 mL medium intraperitoneally, wash out after 4 hours.