Chapter title |
Southwestern Blotting Assay
|
---|---|
Chapter number | 5 |
Book title |
DNA-Protein Interactions
|
Published in |
Methods in molecular biology, January 2015
|
DOI | 10.1007/978-1-4939-2877-4_5 |
Pubmed ID | |
Book ISBNs |
978-1-4939-2876-7, 978-1-4939-2877-4
|
Authors |
Yinshan Jia, Linda Nagore, Harry Jarrett, Jia, Yinshan, Nagore, Linda, Jarrett, Harry |
Abstract |
Southwestern blotting is a technique used to study DNA-protein interactions. This method detects specific DNA-binding proteins by incubating radiolabeled DNA with a gel blot, washing, and visualizing through autoradiography. A blot resulting from 1-dimensional SDS-PAGE reveals the molecular weight of the binding proteins. To increase separation and determine isoelectric point a 2-dimensional gel can be blotted. Additional dimensions of electrophoresis, such as a gel shift (EMSA), can precede isoelectric focusing and SDS-PAGE to further improve separation. Combined with other techniques, such as mass spectrometry, the DNA-binding protein can be identified. |
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