This is the first report on the determination of proteins with 3-hydroxy-4-(2-hydroxy-4-sulfo-1-naphthalenyl)azo (Cal-Red) by Rayleigh light scattering (RLS). At pH 4.07, the weak RLS of Cal-Red can be enhanced greatly by the addition of proteins. On this basis, the reaction of Cal-Red and proteins was studied. A new quantitative determination method for proteins has been developed. This method is very sensitive (0.45-36.9 microgml(-1) for bovine serum albumen (BSA)), rapid (<2min), simple (one step), and tolerant of most interfering substances. The maximum binding number of Cal-Red to BSA was 143 and the binding constant was 4.1x10(6)mol(-1)L. Four samples of total protein in human serum were determined and the maximum relative error is no more than 3%.