I'm looking for a simple UV-vis based protein ligand binding assay that a high school level or beginner college level student can perform independently. As such it would be best if it had step by step procedure and calculations. Thank you in advance.
UV-VIS-based measurement of enzyme catalysis with a substrate is very common and easy. I can't think of an example of measuring binding of a ligand to a protein by UV-VIS without a subsequent chemical reaction. Generally speaking, there is not a substantial difference between the spectrum that is the sum of the spectra of the 2 binding partners and the spectrum of the complex.
Some examples of photometric measurements of enzyme reactions with chromogenic substrates: beta-galactosidase with ortho-nitrophenylgalactoside (ONPG), beta-lactamase with nitrocefin, horseradish peroxidase with hydrogen peroxide and tetramethylbenzidine (TMB), and alkaline phosphatase with para-nitrophenylphosphate.
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