Phospholipase A2 (PLA2) assay

The PLA₂ activity of the venom samples was assessed via turbidimetric assay. The substrate for the reaction was freshly prepared with chicken egg yolk dissolved in 0.9% NaCl solution, and its absorbance was made up to one at 740 nm [20,21]. The time-dependent kinetic assay was performed in triplicate with different concentrations of each venom sample (0.01 μg, 0.1 μg, 0.5 μg, and 1 μg) prepared in a 20 mM Tris-Cl buffer. The resulting absorbance was measured at 1-minute intervals for 60 mins at 740 nm using an Epoch 2 microplate spectrophotometer (BioTek Instruments, Inc., USA). Unit activity was calculated as the amount of crude venom required to reduce the absorbance of the substrate by 0.01 OD unit per min at the given wavelength [22].