A NEW METHOD TO ASSIGN IMMUNODETECTED SPOTS IN THE COMPLEX 2-DIMENSIONAL ELECTROPHORESIS PATTERN

A new, easy method for the immunodetection of specific antigens in two -dimensional electrophoresis (2-DE) is described. Areas of 2-DE gels c ontaining antigens of interest are electrophoretically transferred to polyvinylidene difluoride membranes, immunostained with specific antib odies using Fast Red or 5-bromo-4-chloro-3-indolyl phosphate/nitro blu e tetrazolium as detection systems and counterstained with Coomassie B rilliant Blue. In contrast to conventional methods, it is possible to use this procedure to exactly assign immunoreacting proteins on a sing le blot to their corresponding and surrounding blue-stained protein sp ots.