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Updated: Jun 23, 2026

Quantitative Detection of DNA-Protein Crosslinks and Their Post-Translational Modifications
Published on: April 21, 2023
Identification of sequence-specific DNA-binding proteins by southwestern blotting
Simon Labbé1, Jean-François Harrisson, Carl Séguin
1Centre de Recherche en Cancérologie de l'Université Laval, CHUQ, Pavillon l'Hôtel-Dieu de Québec, 11, côte de Palais, Québec, QC, G1R 2J6, Canada.
Abstract:
We describe a Southwestern blotting method for characterization of both DNA-binding proteins and their specific sites. Proteins are first separated on a sodium dodecyl sulfate (SDS) polyacrylamide gel, then renatured in SDS-free buffer and transferred by electroblotting to an immobilizing membrane, and detected by their ability to bind radiolabeled DNA. The protein(s) interacting with the labeled DNA is visualized by autoradiography. This technique was used in our laboratory to visualize the metal regulatory consensus sequence-binding protein MTF-1 in L cell crude nuclear extracts.
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Southern Blot
Denatured DNA fragments must be transferred onto a carrier membrane from the gel to make it accessible to a probe - a small ssDNA fragment complementary to the target DNA...
Western Blotting
The technique begins with separating proteins from the sample using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), followed by protein transfer, immunoblotting, and finally, protein detection.
Single-Strand DNA Binding Proteins

