Chapter title |
Simultaneous Immunoblotting Analysis with Activity Gel Electrophoresis and 2-D Gel Electrophoresis
|
---|---|
Chapter number | 9 |
Book title |
Western Blotting
|
Published in |
Methods in molecular biology, January 2015
|
DOI | 10.1007/978-1-4939-2694-7_9 |
Pubmed ID | |
Book ISBNs |
978-1-4939-2693-0, 978-1-4939-2694-7
|
Authors |
Der-Yen Lee, Geen-Dong Chang |
Editors |
Biji T. Kurien, R. Hal Scofield |
Abstract |
Diffusion blotting method can couple immunoblotting analysis with another biochemical technique in a single polyacrylamide gel, however, with lower transfer efficiency as compared to the conventional electroblotting method. Thus, with diffusion blotting, protein blots can be obtained from an SDS polyacrylamide gel for zymography assay, from a native polyacrylamide gel for electrophoretic mobility shift assay (EMSA) or from a 2-D polyacrylamide gel for large-scale screening and identification of a protein marker. Thereafter, a particular signal in zymography, electrophoretic mobility shift assay, and 2-dimensional gel can be confirmed or identified by simultaneous immunoblotting analysis with a corresponding antiserum. These advantages make diffusion blotting desirable when partial loss of transfer efficiency can be tolerated or be compensated by a more sensitive immunodetection reaction using enhanced chemiluminescence detection. |
Mendeley readers
Geographical breakdown
Country | Count | As % |
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Unknown | 12 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
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Student > Bachelor | 3 | 25% |
Student > Ph. D. Student | 3 | 25% |
Researcher | 2 | 17% |
Professor | 1 | 8% |
Student > Master | 1 | 8% |
Other | 1 | 8% |
Unknown | 1 | 8% |
Readers by discipline | Count | As % |
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Agricultural and Biological Sciences | 7 | 58% |
Biochemistry, Genetics and Molecular Biology | 3 | 25% |
Unknown | 2 | 17% |