Myelotracker allows one-step staining of neutrophils. It is supplied at a 1 mg/mL concentration and should be used like any other fluorescent marker (ie. Dapi, fluorescent Phalloidin, …).
Fixed samples. We recommend to use Myelotracker at a 1 microg/mL concentration in PBS with or without a permeabilizer, depending on the application. 1 hour-straining at room temperature is sufficient for efficient labelling of neutrophil granules or secretion.
Live cells. Myelotracker+ (proteolysis-resistant) is recommended to assess neutrophil degranulation in vitro. It may be incubated with neutrophils in culture (RPMI 1640 medium supplemented with 10mM Hepes and 10% inactivated human serum, or similar medium). We recommend to use Myelotracker at 1 microg/mL to assess neutrophil degranulation.
Intravital imaging (animal models). Neutrophil degranulation may be assessed in animal models of inflammation or infection, upon i.v. injection of Myelotracker+ (proteolysis-resistant). We recommend to inject 5 microg Myelotracker+/mouse. Myelotracker+ detection may be recorded for at least 48 hours.