The combination of Indian ink staining with immunochemiluminescence detection allows precise identification of antigens on blots: application to the study of glycosylated barley storage proteins.
The localization after blotting of specific spots in two-dimensional electrophoretic protein pattern was achieved using, in that order, Indian ink protein staining and immunodetection with chemiluminescence on the same membrane. Indian ink did not inhibit significantly the antibody reactions even after overnight staining. It produces permanent staining that did not quench the chemiluminescent signal, recorded on a film. This allowed perfect matching between the specific and the total protein patterns. The procedure was applied to the identification of glycoproteins present in barley storage protein preparations.
Eynard
L
L
Laboratoire de Chimie Biologique, Centre INRA de Grignon, Thiverval-Grignon, France. eynard@cardere.grignon.inra.fr
Laurière
M
M
D000941:Antigens / Q000032:analysis; D002244:Carbon; D004396:Coloring Agents; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D005983:Glutens; D006023:Glycoproteins / Q000032:analysis*; D006031:Glycosylation; D001467:Hordeum / Q000378:metabolism*; D015151:Immunoblotting / Q000379:methods*; D008163:Luminescent Measurements; D010940:Plant Proteins / Q000032:analysis*; D013194:Staining and Labeling / Q000379:methods
D000941:Antigens; D004396:Coloring Agents; D006023:Glycoproteins; D010940:Plant Proteins; C028433:chinese ink; D002244:Carbon; D005983:Glutens