I am using one enzyme to find out the specific activity of different substrates at different wavelengths. Is this right? or should I need to use same wavelength for all substrates for a single enzyme?
You should use whatever wavelength is appropriate for each substrate. You will need a standard curve or relevant extinction coefficient for each substrate at the wavelength used for the measurement in order to convert absorbance (or fluorescence) change to concentration.
You should use whatever wavelength is appropriate for each substrate. You will need a standard curve or relevant extinction coefficient for each substrate at the wavelength used for the measurement in order to convert absorbance (or fluorescence) change to concentration.