Bovine Serum Albumin Synthesis Lab Report

2.5. Bovine Serum Albumin (BSA) binding Studies:
BSA solution was prepared in 5 mM Tris-HCl/10 mM NaCl buffer solution to keep pH value constant (pH=7.4) and it is stored at 4oC, used within 2h. The BSA concentration was estimated spectrophotometrically (ε280=44,300 M-1cm-1) [35], 60 µM BSA was titrated with increasing concentrations of Ru (II) complex (0-50 µM). Absorbance changes at 280 nm were monitored by UV-spectrometry.

2.6. DNA Binding Studies:
Binding studies of DNA were carried out at room temperature. The tris buffer (5 mM Tris-HCl, 50 mM NaCl, pH=7.1) was prepared using doubly distilled water. At fixed concentration of Ru(II) complexes(10 μM) in the buffer, by increasing the DNA concentration absorption spectra were recorded. The binding strength of the complex

[DNA]/(Ԑa-Ԑf) versus [DNA] graph was plotted. Slope of the straight line gives Kb

All the complexes were tested for their antimicrobial activity against viz. E.coli, Bacillus subtilis and Staphylococcus aureus. Two different concentrations, 1mg/ml (1000 μM) and 0.5 mg/ml (500 μM) in DMSO were used for testing spore germination of each fungus. 5 mm size filter paper disks were prepared using whatmann filter paper no.1 saturated with 10 μl of complexes and placed in petri dishes containing LB (Luria Bertini) agar media inoculated with E. coli, Bacillus subtilis and Staphylococcus aureus separately. The petri dishes were incubated at 25±0.2°C and the zone of inhibitions were noticed after 48h. The plates were observed and the diameters of the inhibition zones (in mm) were deliberated, and compared with standard antibacterial drug