Neutrophil granulocytes are essential players in the inflammatory response. Many functional processes in neutrophils, including migration depend on changes in cytosolic free Ca²⁺ concentration ([Ca²⁺]_c). Neutrophils respond to a range of chemotactic factors such as N-formyl-methionyl-leucyl-phenylalanine (fMLP) and interleukin-8 (IL-8), triggering Ca²⁺ release from intracellular stores via opening of inositol-trisphosphate (IP₃) receptors. The Golgi apparatus is an intracellular Ca²⁺ store, which accumulates Ca²⁺ via both thapsigargin-sensitive sarco-endoplasmic reticulum Ca²⁺ ATPases (SERCAs) and the thapsigargin-insensitive secretory pathway Ca²⁺ transport ATPases (SPCAs). Until now, IP₃-producing agonists have been reported to mobilize only the thapsigargin-sensitive part of the Golgi Ca²⁺ store. In this study, agonist-induced Ca²⁺ -release from the SERCA-independent intracellular stores was measured in the presence of the SERCA-specific inhibitor thapsigargin. fMLP and IL-8 were applied as agonists. Fura-2 was used as Ca²⁺ probe. fMLP mobilized the SERCA-independent store in 40% of the neutrophil granulocytes, while IL-8 induced Ca²⁺ release from SERCA-independent store in 20% of the neutrophils. The expressions of both human SPCA isoforms, hSPCA1 and hSPCA2 in neutrophils was shown by Western blotting and immunocytochemistry. One third of the neutrophil granulocytes retained migratory activity in the presence of the SERCA inhibitor thapsigargin in Ca²⁺ -free solution, while in the presence of the intracellular Ca²⁺ chelator BAPTA, the migration was nearly abolished. These results suggest that a SERCA-independent and probably SPCA-dependent intracellular Ca²⁺ store might be important in Ca²⁺ signaling and migration of human neutrophil granulocytes.