Sodium dodecyl sulphate-polyacrylamide denaturing gel electrophoresis and Western blotting techniques.
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.
Wellcome Trust Centre for Human Genetics, Headington, Oxford, UK. christin@well.ox.ac.uk
Christine
C
Blancher
Rob Mc
RM
Cormick
D001391:Azo Compounds / Q000378:metabolism; D015153:Blotting, Western / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D008567:Membranes, Artificial; D010455:Peptides / Q000378:metabolism; D011506:Proteins / Q000302:isolation & purification* / Q000378:metabolism; D012394:Rosaniline Dyes / Q000378:metabolism; D013194:Staining and Labeling
D001391:Azo Compounds; D008567:Membranes, Artificial; D010455:Peptides; D011506:Proteins; D012394:Rosaniline Dyes; C004692:coomassie Brilliant Blue; C032756:ponceau S