Activity and release of myeloperoxidase (MPO) was measured in heparinized whole blood samples after activation of neutrophil granulocytes by the chemoattractant N-formyl-methionyl-leucyl-phenylalanine (fMLP) using two different methods: (i) by determination of the amount of MPO released into the blood plasma using a MPO enzyme-immunoassay, and (ii) simultaneously, by measuring the remaining activity within the neutrophils by flow cytometry using the Bayer Technicon H3. Although a part of MPO was released immediately after addition of fMLP, remaining MPO activity within the neutrophils surprisingly increased during the first minutes after incubation. Subsequently, MPO activity dropped due to a continuous release of MPO. In addition to fMLP, granulocyte-macrophage colony stimulating factor (GM-CSF) enhanced MPO activity in neutrophils. These results indicate that MPO is present in resting granulocytes in an inactive or only partially active form and is activated by fMLP and GM-CSF.