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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Kurien, B.T., Danda, D., Bachmann, M.P., Scofield, R.H. (2015). SDS-PAGE to Immunoblot in One Hour. In: Kurien, B., Scofield, R. (eds) Western Blotting. Methods in Molecular Biology, vol 1312. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2694-7_45
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DOI: https://doi.org/10.1007/978-1-4939-2694-7_45
Publisher Name: Humana Press, New York, NY
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