In the subcellular isolation experiment, sucrose and mannitol is present in buffers for resuspending cell pellet after centrifugation. For example, Isolation buffer (IB): 220 mM d-mannitol, 70 mM d-sucrose, 5 mM HEPES-KOH pH 7.4, 1 mM EGTA-KOH pH 7.4. And then cells are lysed by pressurizing nitrogen.