AN INVESTIGATION OF THE FACTORS CONTROLLING THE STAINING AND DESTAINING OF ELECTROPHORESIS GELS

A general analysis is developed for the effect of mass transport and k inetic parameters on the rate of staining and destaining of electropho resis gels. The various contributions to these overall processes are d iscussed and, in particular, the ways in which the rate of solution fl ow, temperature and gel properties can influence staining procedures a re highlighted. It is shown that for reproducible, rapid and uniform s taining a rotating gel system, analogous to a rotating disc, provides the necessary controlled laminar flow. The theoretical equations deriv ed are compared with experiments and it is shown that at high gel rota tion speeds mass transport in solution does not have a major controlli ng influence on rates of staining and destaining. The temperature depe ndence of these rates also suggests that there is no significant contr ol by the rate of interaction between stain and protein molecules. The major controlling factor under these conditions is then concluded to be the transport of stain in the gel itself, and the theoretical analy sis and time-dependent experimental results allow a determination of t he corresponding diffusion parameters.