Immunodetection after complete destaining of coomassie blue-stained proteins on immobilon-PVDF.
A technique that simplifies the localization of an immunodetectable protein in relation to the other electrophoresed proteins is described. Proteins are transblotted onto a polyvinylidene difluoride (PVDF) membrane and visualized by staining with Coomassie brilliant blue R-250, and a photograph of the protein pattern is taken. The Coomassie blue-stained PVDF membrane is then completely destained using a 25% acetic acid/50% methanol solution that allows subsequent immunostaining on the same membrane. The technique uses common laboratory reagents, is rapid, and has been shown to be applicable for a variety of proteins using both monoclonal and polyclonal antibodies and a variety of transblots.
Pryor
J L
JL
Department of Cell Biology, University of Minnesota, Minneapolis 55455.
Xu
W
W
Hamilton
D W
DW
D000818:Animals; D000906:Antibodies; D000911:Antibodies, Monoclonal; D004591:Electrophoresis, Polyacrylamide Gel; D004822:Epididymis / Q000737:chemistry*; D007124:Immunoenzyme Techniques; D008297:Male; D008567:Membranes, Artificial; D011145:Polyvinyls; D011506:Proteins / Q000032:analysis*; D051381:Rats; D011919:Rats, Inbred Strains; D012394:Rosaniline Dyes / Q000737:chemistry; D013194:Staining and Labeling; D005465:Fluorocarbon Polymers
D000906:Antibodies; D000911:Antibodies, Monoclonal; D008567:Membranes, Artificial; D011145:Polyvinyls; D011506:Proteins; D012394:Rosaniline Dyes; C024865:polyvinylidene fluoride; D005465:Fluorocarbon Polymers; C048139:Coomassie blue