Skip to main content
. Author manuscript; available in PMC: 2021 May 4.
Published in final edited form as: Methods Mol Biol. 2021;2251:225–236. doi: 10.1007/978-1-0716-1142-5_17

Fig. 3.

Fig. 3

Size of lipid vesicle affects the relative rate of XY-69 hydrolysis by PLC-γ1 (WT) and PLC-γ1 (D1165H). (a, b) The Z-average (Zav) particle size of lipid vesicles was measured by dynamic light scattering (DLS). The % intensity of total scattered light was plotted versus particle size of lipid vesicles, which were formed either by probe sonication only (a) or by first probe sonication and then freeze–thaw (b). (c, d) XY-69 (0.5 μM) hydrolysis by PLC-γ1 (WT) or PLC-γ1 (D1165H) at 20 °C was monitored by fluorescence (λex/λem = 485/520 nm). Lipid vesicles were formed either by probe sonication only (c) or by first probe sonication and then freeze–thaw (d). All lipid vesicles contain PE (192 μM) and PIP2 (48 μM). The final pH of the assay buffer was 7.4, and the free Ca2+ concentration was 390 nM