Chapter title |
SDS-PAGE to Immunoblot in One Hour
|
---|---|
Chapter number | 45 |
Book title |
Western Blotting
|
Published in |
Methods in molecular biology, January 2015
|
DOI | 10.1007/978-1-4939-2694-7_45 |
Pubmed ID | |
Book ISBNs |
978-1-4939-2693-0, 978-1-4939-2694-7
|
Authors |
Biji T. Kurien Ph.D., Debashish Danda M.D., Michael P. Bachmann Ph.D., R. Hal Scofield M.D., Biji T. Kurien, Debashish Danda, Michael P. Bachmann, R. Hal Scofield |
Editors |
Biji T. Kurien, R. Hal Scofield |
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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