Celltiter 96 aqueous non radioactive cell proliferation assay kit mts

Cells were collected using accutase (Innovative cell technologies, San diego, CA) and then stained with trypan blue solution. Viable cells, trypan blue negative, were counted using hemocytometer under microscope. Proliferation was determined with celltiter 96 aqueous nonradioactive cell proliferation assay kit (MTS; 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxyme-thoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, Promega, Madison, WI, USA) as described in the manufacturer's instruction. Absorbance was measured at 490 nm with a powerwave HT spectrophotometer (Biotek instruments, Winooski, VT, USA).

Prosopilosidine (1), isoprosopilosidine (2), prosopilosine (4), isoprosopilosine (5), and juliprosopine (6) were isolated from P. glandulosa (Fam. Leguminosae), which was collected from Nevada, USA, as described previously by Samoylenko et al. (2009) [29 ]. In addition, compounds (3) and (6) were isolated from the P. glandulosa sample that was collected in Texas. Amp B and Flu were purchased from Sigma-Aldrich (St. Louis, MO, USA). Amphotericin B and PPD were separately dissolved as a stock solution (1 mg/mL containing 13.3 µL DMSO, 13.3 µL Tween-20, and 973.4 µL distilled water) as colored and clear solutions, respectively. They were protected from light and were stored in the refrigerator. Different doses were prepared from this stock and were used daily. CellTiter 96^® AQueous Non-Radioactive Cell Proliferation Assay kit (MTS) was purchased from Promega (Madison, WI, USA).