The relative fluorescence intensity of ANS-protein conjugates was measured using an PerkinElmer LS45
spectrofluorometer (American PerkinElmer Inc.) at excitation wavelength 365 nm and emission wavelength 480 nm.
After chromogen staining, the fluorescence of each well was measured in a
spectrofluorometer (Thermo Scientific Lumina, USA) at 450 nm.
The implementation of fluorescence measurements was with the help of RF-5301PC
spectrofluorometer (Shimadzu, Japan), and the slit width was 5.0 nm.
UV-vis spectra were obtained with a PerkinElmer Lambda 356 UV/VIS spectrophotometer, and emission measurements were performed on a Fluorolog-3
spectrofluorometer. [sup.1]H and [sup.13]C spectra were recorded on a Bruker avance DPX 400.
Emission and excitation spectra were measured on QuantaMaster 40 steady state
spectrofluorometer (Photon Technology International, Inc.).
The fluorescence spectra were obtained using an FP-6500
spectrofluorometer (JASCO Co., Tokyo, Japan).
The PerkinElmer model LS 55 was employed throughout this experiment; however, any other commonly available
spectrofluorometer (Horiba, Hitachi, or Shimatsu), with dual gratings, one for excitation wavelength selection and another for emission band scanning, will be equally suitable.