Chapter title |
Diffusion Blotting: A Rapid and Simple Method for Production of Multiple Blots from a Single Gel
|
---|---|
Chapter number | 10 |
Book title |
Western Blotting
|
Published in |
Methods in molecular biology, January 2015
|
DOI | 10.1007/978-1-4939-2694-7_10 |
Pubmed ID | |
Book ISBNs |
978-1-4939-2693-0, 978-1-4939-2694-7
|
Authors |
Ingrid Olsen D.V.M., Ph.D., Harald G. Wiker M.D., Ph.D., Ingrid Olsen, Harald G. Wiker |
Editors |
Biji T. Kurien, R. Hal Scofield |
Abstract |
A very simple and fast method for diffusion blotting of proteins from precast SDS-PAGE gels on a solid plastic support was developed. Diffusion blotting for 3 min gives a quantitative transfer of 10 % compared to 1-h electroblotting. For each subsequent blot from the same gel a doubling of transfer time is necessary to obtain the same amount of protein onto each blot. High- and low-molecular-weight components are transferred equally efficiently when compared to electroblotting. However, both methods do give a higher total transfer of the low-molecular-weight proteins compared to the large proteins. The greatest advantage of diffusion blotting is that several blots can be made from each lane, thus enabling testing of multiple antisera on virtually identical blots. The gel remains on the plastic support, which prevents it from stretching or shrinking. This ensures identical blots and facilitates more reliable molecular weight determination. Furthermore the proteins remaining in the gel can be stained with Coomassie Brilliant Blue or other methods for exact and easy comparison with the developed blots. These advantages make diffusion blotting the method of choice when quantitative protein transfer is not required. |
Mendeley readers
Geographical breakdown
Country | Count | As % |
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Unknown | 4 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
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Student > Ph. D. Student | 1 | 25% |
Student > Bachelor | 1 | 25% |
Researcher | 1 | 25% |
Student > Postgraduate | 1 | 25% |
Readers by discipline | Count | As % |
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Biochemistry, Genetics and Molecular Biology | 2 | 50% |
Pharmacology, Toxicology and Pharmaceutical Science | 1 | 25% |
Agricultural and Biological Sciences | 1 | 25% |