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Fluorescence emission spectra of EnzChek® Phospholipase A2 substrate incorporated in liposomes with addition
of bee venom PLA2 at ambient temperature.
Plot of fluorescence emission intensities versus concentration of PLA2 per well at 10 minutes, run at ambient temperature with liposomes. Fluorescence was measured exciting at 460 nm on a Spectra Max M5 (Molecular Devices).
Panel A: Ratiometric (FRET)-based (515/575 nm) EnzChek® Phospholipase A2 substrate. Panel B: Intensity-based detection
mode (515 channel)- EnzChek® Phospholipase A2 substrate. Background fluorescence determined for the no-PLA2 enzyme
control reaction has been subtracted for each value.
IC50 determination for Manoalide, a known inhibitor1 using EnzCheck® Phospholipase A2 Assay Kit. Samples
of Manoalide were titrated in the presence of 1.65 × 10-6 M Red/Green BODIPY® PC-A2 and 0.125 U/mL PLA2 from bee
venom in 1X PLA2 reaction buffer. Reactions were incubated for 5 minutes at room temperature and fluorescence was
measured at Ex/Em 460/515 nm using a microplate reader.