A nonradioactive double detection method for the assignment of spots in two-dimensional blots.
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.
Chevallet
M
M
CEA-Laboratoire de Bioénergétique Cellulaire et Pathologique, Grenoble, France.
Procaccio
V
V
Rabilloud
T
T
D001426:Bacterial Proteins / Q000302:isolation & purification; D005069:Evaluation Studies as Topic; D017979:Gold Colloid; D015151:Immunoblotting / Q000379:methods*; D008163:Luminescent Measurements; D008565:Membrane Proteins / Q000302:isolation & purification; D011506:Proteins / Q000302:isolation & purification*; D016354:Rhodobacter capsulatus / Q000737:chemistry; D012834:Silver; D013194:Staining and Labeling
D001426:Bacterial Proteins; D017979:Gold Colloid; D008565:Membrane Proteins; D011506:Proteins; D012834:Silver