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initiation of the apoptotic pathway.

To find the inhibitory effect of STS on calcium oxalate stone formation the gel diffusion assay was performed according to Li et al. with minor modifications A microscope slide was uniformly coated with 3mL of 1% agar. After the agar solidified, two pairs of equidistant wells were made perpendicularly. Sodium oxalate and calcium chloride each 20μl was placed in vertical wells. The horizontal wells were filled either with 20μl distilled water as standard or 20μl of STS at varying concentrations. Then the slide was left in a moist chamber for 24h at room temperature. The calcium and oxalate ions diffuse through the gel and form crystals of calcium oxalate, visible as a cloudy streak in the center. The intensity of crystal formation and size of the crystals was dependent on the molarity of the crystal forming solutions employed. Depending on concentration, the inhibitory substances would modify the density and width of the crystal streak. This was carried out in triplicates with different concentrations (200mM, 100mM, 50mM, and 25mM) of STS. The slides were photographed using Gel Documentation System ‘BioRad Chemidoc XRS’. The images were analyzed using Image J software and densitometry plots were obtained. Relative density of the sample with respect to the control was obtained, and the percentage inhibition was calculated by the following formula: inhibition=1-(relative density of the sample/relative density of the control)*100.