Separation of chlorophyll‐protein complexes by deriphat polyacrylamide gradient gel electrophoresis

Abstract

An improved Deriphat polyacrylamide gradient gel electrophoresis system was developed for the separation of chlorophyll‐protein complexes. The relatively good resolution of the starting discontinuous gel system was further improved by using glycerol in gels and an acrylamide gradient with high acrylamide‐to‐N,N′,‐methylenebisacrylamide ratio in the separating gel. By applying mild but efficient glycosidic detergents for solubilization, and Deriphat to gels and buffers, the stability of complexes was increased, and only a low amount of pigment was removed. The advantage of our system is the better resolution of larger‐size complexes, especially those of photosystem I. In addition, it makes possible an easier interpretation of results due to less overlapping of photosystem I and photosystem II bands when different plant species or the effects of different treatments are compared using whole thylakoid membranes.