Supplementary MaterialsFigure S1: Optical photograph depicting change in color of reaction
May 10, 2019
Supplementary MaterialsFigure S1: Optical photograph depicting change in color of reaction mixture as a result of change in surface plasmon resonance upon incubation of HAuCl4 solution (10?3 M) with increasing amounts of dithiothreitol. analysis, and atomic force microscopy revealed the LY2228820 manufacturer size of spherical gold nanoparticles to be in the range of 20C40 nm and nonspherical gold particles were found to be 60C80 nm. We also evaluated the potential of biogenic gold nanoparticles to probe liver cancer cells by conjugating them with liver cancer cell surface-specific antibodies. The antibody-conjugated gold particles were found to bind specifically to the surface antigens of the cancer cells. Conclusion The antibody-conjugated gold particles synthesized in this study could successfully differentiate normal cell populations from cancerous cells. was cultured on YEDP agar plates. The cells were harvested after 24 hours and homogenized in chilled lysis buffer supplemented with a protease inhibitor cocktail, ie, 2% Triton X-100 (w/v), 1% sodium OBSCN dodecyl sulfate, 100 mM Tris-HCl (pH 8.0), 100 mM NaCl, 1 mM ethylenediamine tetra-acetic acid (pH 8.0), and 1 mM phenylmethylsulfonyl fluoride. The homogenate was sonicated for 45 minutes at 4C using a bath sonicator. Subsequently, the homogenate was vortexed for 1 hour with intermittent cooling at 4C. The preparation was pelleted at 2000 g for 15 minutes, and the LY2228820 manufacturer supernatant was collected and kept at ?20C until further use. Synthesis of gold nanoparticles using cytosolic extract Different volumes (1C5 mL) of cytosolic extract were added to 5 mL solution of 10?3 M aqueous HAuCl4, and the volume was made up to 10 mL by adding the appropriate amount of deionized water. The mixture was incubated for 24 hours to complete the reaction. The synthesized gold nanoparticle product was characterized by ultraviolet-visible spectroscopy, transmission electron microscopy, atomic force microscopy, and Fourier transform infrared analyses. Ultraviolet-visible and fluorescence spectroscopy To characterize the synthesized gold nanoparticles, they were scanned in the 300C1000 nm wavelength range using a double beam spectrophotometer (Perkin Elmer, Boston, MA). Fluorescence spectra were recorded with a Hitachi F-4500X fluorescence spectrometer (Hitachi, Tokyo, Japan) controlled by a personal computer data processing unit. The excitation at 488 nm and the emission spectra from 505 nm to 550 nm were collected. All excitation and emission slits were set at 5 nm. Transmission electron microscopy The size, shape, and morphology of the synthesized gold nanoparticles were analyzed using a transmission electron microscope (1200 EX, JOEL Inc, Peabody, MA) following a method described elsewhere.26 Samples were prepared by putting a drop of the gold particles on a negative carboncoated copper grid and dried in air before being transferred to the transmission electron microscope. Fourier transform infrared spectroscopy Fourier transform infrared spectroscopic measurement of the gold nanoparticles was carried out by depositing purified particles on Si (111) wafers with simple dropcoating and subjecting them to analysis (6700 spectrum; Thermo Nicolet, Madison, WI) in a diffuse reflectance mode at a resolution of 4 cm?1. Atomic force microscopy Samples of the biogenic gold nanoparticles generated using cytosolic extract were centrifuged and redispersed in deionized water. Samples were passed through a 0.22 m filter, and an aliquot of reaction mixture was LY2228820 manufacturer placed on a Si disc and dried in a nitrogen atmosphere. LY2228820 manufacturer Finally, the samples were analyzed using a contact mode atomic force microscope (Perkin Elmer). Animals Female BALB/c mice weighing 18 2 g and aged 8C10 weeks were obtained from the animal house facility of the Institute of Microbial Technology and used for induction of.