Sodium dodecyl sulphate-polyacrylamide denaturing gel electrophoresis and Western blotting techniques

• Published in: Methods in molecular biology (Clifton, N.J.) Vol. 878; p. 89
• Main Authors: Blancher, Christine, Cormick, Rob Mc
• Format: Journal Article
• Language: English
• Published: United States 2012
• Subjects: Azo Compounds - metabolism; Blotting, Western - methods; Electrophoresis, Polyacrylamide Gel - methods; Membranes, Artificial; Peptides - metabolism; Proteins - isolation & purification; Proteins - metabolism; Rosaniline Dyes - metabolism; Staining and Labeling
• ISSN: 1940-6029
• DOI: 10.1007/978-1-61779-854-2_6

The study of proteins, their expression and post-translational modification, is a key process in molecular biology. Immunoblotting is a well-established and powerful tool for the study of proteins, which continues to evolve as new reagents and apparatus are developed. This chapter describes in detail the process by which proteins are extracted from cells, quantified, fractionated using poly-acrylamide gel electrophoresis, transferred to a membrane, and assessed by immunoblotting. Variations in experimental technique, and new technologies available to the researcher, are also discussed.