Chapter title |
Western Blotting Using PVDF Membranes and Its Downstream Applications
|
---|---|
Chapter number | 24 |
Book title |
Western Blotting
|
Published in |
Methods in molecular biology, January 2015
|
DOI | 10.1007/978-1-4939-2694-7_24 |
Pubmed ID | |
Book ISBNs |
978-1-4939-2693-0, 978-1-4939-2694-7
|
Authors |
Setsuko Komatsu Ph.D., Setsuko Komatsu |
Editors |
Biji T. Kurien, R. Hal Scofield |
Abstract |
Western blotting using polyvinylidene difluoride (PVDF) membranes is one of the most popular techniques for detection and characterization of proteins. If this technique is combined with immunodetection, the behavior of a particular protein can be clarified. On the other hand, if it is combined with Edman sequencing, the primary structure of the protein can be determined. A protein sample is transferred from an SDS-polyacrylamide gel electrophoresis (PAGE) gel onto a PVDF membrane by electroblotting. The membrane carrying the protein is either used for immunodetection or protein sequencing. SDS-PAGE followed by Western blotting combined with immunodetection using antibodies can easily detect protein behavior in crude protein mixtures. Furthermore, two-dimensional PAGE followed by Western blotting and Edman sequencing allows effective sequence determination of crude protein mixtures that may not be easily purified by conventional column chromatography. |
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