SDS-PAGE to Immunoblot in One Hour
Biji T Kurien1, Debashish Danda, Michael P Bachmann
1Arthritis and Clinical Immunology Program, Oklahoma Medical Research Foundation, 825 NE 13th Street, Oklahoma City, OK, 73104, USA, biji-kurien@omrf.org.
Abstract:
An ultra-rapid method for electrophoresing proteins on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, transfer of proteins to nitrocellulose membranes, and immunoblotting is described here. Electrophoresis of the autoantigens La and Ro60, as well as molecular weight standards on a 4-20 % gradient gel, was performed in about 10 min using heated (70-75 °C) normal Laemmli running buffer. Electrophoretic transfer of these proteins was achieved in 7 min using a semidry transfer method. Finally, immunoblotting of La and Ro60 was carried out in 30 min. Thus, the entire process of electrophoresis, electrotransfer, and immunoblotting could be carried out in 1 h.
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