Western blotting

• Published in: Methods (San Diego, Calif.) Vol. 38; no. 4; pp. 283 - 293
• Main Authors: Kurien, Biji T., Scofield, R. Hal
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01-04-2006
• Subjects: Animals; Antigens - chemistry; Biotinylation; Blotting, Western - instrumentation; Blotting, Western - methods; Collodion - chemistry; Detection techniques; Diffusion; Electrophoresis; Electrophoresis, Polyacrylamide Gel; Humans; Immunoblotting; Membranes, Artificial; Nitrocellulose; Polyvinylidene difluoride membrane; Proteins - analysis; Proteins - chemistry; Sodium dodecyl sulfate–polyacrylamide gel electrophoresis; Solvents; Western blotting; Polyvinylidene difluoride membrane; Sodium dodecyl sulfate–polyacrylamide gel electrophoresis; Western blotting; Nitrocellulose; Detection techniques
• ISSN: 1046-2023; 1095-9130
• DOI: 10.1016/j.ymeth.2005.11.007

Western blotting (protein blotting or immunoblotting) is a powerful and important procedure for the immunodetection of proteins post-electrophoresis, particularly proteins that are of low abundance. Since the inception of the protocol for protein transfer from an electrophoresed gel to a membrane in 1979, protein blotting has evolved greatly. The scientific community is now confronted with a variety of ways and means to carry out this transfer. This review describes the various procedures that have been used to transfer proteins from a gel to a membrane based on the principles of simple diffusion, vacuum-assisted solvent flow and electrophoretic elution. Finally, a brief description of methods generally used to detect antigens on blots is also described.