A new method to assign immunodetected spots in the complex two-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 containing antigens of interest are electrophoretically transferred to polyvinylidene difluoride membranes, immunostained with specific antibodies using Fast Red or 5-bromo-4-chloro-3-indolyl phosphate/nitro blue tetrazolium as detection systems and counterstained with Coomassie Brilliant Blue. In contrast to conventional methods, it is possible to use this procedure to exactly assign immunoreacting proteins on a single blot to their corresponding and surrounding blue-stained protein spots.
Zeindl-Eberhart
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Institute of Pathology, University of Munich, Germany.
Jungblut
P R
PR
Rabes
H M
HM
D000818:Animals; D006528:Carcinoma, Hepatocellular / Q000737:chemistry; D003101:Collodion; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D005982:Glutathione Transferase / Q000032:analysis; D006801:Humans; D015151:Immunoblotting / Q000379:methods*; D007124:Immunoenzyme Techniques; D007202:Indicators and Reagents; D007211:Indoles / Q000737:chemistry; D008297:Male; D008567:Membranes, Artificial; D009580:Nitroblue Tetrazolium / Q000378:metabolism; D011145:Polyvinyls; D011817:Rabbits; D012394:Rosaniline Dyes / Q000737:chemistry; D005465:Fluorocarbon Polymers
D003101:Collodion; D005982:Glutathione Transferase; D007202:Indicators and Reagents; D007211:Indoles; D008567:Membranes, Artificial; D009580:Nitroblue Tetrazolium; D011145:Polyvinyls; D012394:Rosaniline Dyes; C024865:polyvinylidene fluoride; D005465:Fluorocarbon Polymers; C035455:5-bromo-4-chloro-3-indoxyl phosphate; C024757:Coomassie brilliant blue R