Hi everyone, Im completely stuck in interpreting this data. Any help would be greatly appreciated.
Reduced NAD emits fluorescent light at 460 nm when it is excited at 340 nm, and this fluorescence has been found to be enhanced in the presence of lactate dehydrogenase. This effect was used to determine the stoichiometry of binding.
To 2ml of a 0.473 mg/ml solution of beef-heart lactate dehydrogenase in a 0.05M Tris-acetate buffer, pH 7.2, were added small amounts of 10-3 M NADH solution in the same buffer.
The fluorescence emission at 460 nm was measured (in arbitrary units) after each addition and, as a control, readings were also made of the fluorescence changes when the NADH was added to 2ml of the bufferhttp://imgur.com/qrWhDqU
The concentrations of protein in the effluent were estimated by measurements of the absorbance at 280 nm, and the volume of buffer necessary to elute each protein form the column was thus determined.http://imgur.com/5tEaq9o
Thank you so much