A method for identification of spots on two-dimensional electrophoresis gels by means of immunoblotting is described. This method does not require radioactive-labeled proteins and is based on the dual use of colloidal gold staining to detect the general 2D pattern on the blot and on chemiluminescence to detect the antibody-reactive spot(s). Profit is taken from the fact that the general gold stain produces a background pattern on strong ECL exposures, which allows alignment of the antibody-reactive spot(s) on the general pattern and therefore easy identification of the spot(s) of interest on the two-dimensional maps.