Chemicals – Folin ciocalteu reagent, chemicals needed for cell culture and cytotoxicity studies were purchased from Sigma –Aldrich (P.O. box 14 508, St Louis, MO 63178 USA). 1-Diphenyl-2-picrylhydrazyl (DPPH), Triton X-100 solution (1%), gallic acid, sulfanilamide and ortho-phosphoric acid were purchased from Fluka (Fluka chemie GmbH, CH – 9471 Buchs). Tris base was purchased from Promega (Promega Corporation, Madison, WI 53711–5399, USA). All chemicals used were of analytical grade.
Shimadzu UV 1601 UV visible spectrophotometer (Shimadzu Corporation, Kyoto, Japan) was used to measure the absorbance. LFT 600 EC freeze dryer was used to obtain the freeze –dried residue of the AEFLL (LFT 600 EC, −90-95°C temperature, Hitachi pump with 10 valves). Cells were incubated at 37°C in a humidified CO2 incubator (SHEL LAB/Sheldon manufacturing Inc. Cornelius, OR 97113, USA). Olympus (1X70-S1F2) inverted fluorescence microscope (Olympus Optical Co. Ltd. Japan) for observation of cells and photographs were taken using a Nikon D700 camera (Nikon D700, Japan). Deionized water from LABCONCO (waterproplus) UV ultra filtered water system (LABCONCO Corporation, Kansas city, Missouri 64132–2696) or distilled water was used in all experiments.
Shimadzu UV 1601 UV visible spectrophotometer (Shimadzu Corporation, Kyoto, Japan) was used to measure the absorbance. LFT 600 EC freeze dryer was used to obtain the freeze –dried residue of the AEFLL (LFT 600 EC, −90-95°C temperature, Hitachi pump with 10 valves). Cells were incubated at 37°C in a humidified CO2 incubator (SHEL LAB/Sheldon manufacturing Inc. Cornelius, OR 97113, USA). Olympus (1X70-S1F2) inverted fluorescence microscope (Olympus Optical Co. Ltd. Japan) for observation of cells and photographs were taken using a Nikon D700 camera (Nikon D700, Japan). Deionized water from LABCONCO (waterproplus) UV ultra filtered water system (LABCONCO Corporation, Kansas city, Missouri 64132–2696) or distilled water was used in all experiments.