Application of the method of kinetic analysis of staining and destaining processes to the complex formed between hydrolyzed deoxyribonucleoprotein and Schiff's reagent in model films.

Abstract

Recently, we described a method for the analysis of the kinetics of the formation and decomposition of complexes formed between macromolecular substances and covalently bound dyes (Duijndam, Hermans and van Duijn, 1973). Application of this method to the complex formed between the hydrolyzed deoxyribonucleoprotein present in polyacrylamide model films and Schiff's reagent revealed that this complex is formed via a colorless but ultraviolet-absorbing intermediate. Analysis of the process of destaining with 1 N HCl showed that two components are present in the complex, these components differing only slightly in spectral characteristics but significantly in stability toward acid. The main component, making a contribution of about 90% to the total absorbance, was less stable toward acid than the minor (10%) component. The molecular absorbance of the two separate components was found to be 8.2 x 104 and 10.8 x 104 cm2/mmole at 543 nm for the main and minor components, respectively. The conditions that must be checked before conclusions of this kind may be drawn are discussed and controlled.