Western blotting, also known as immunoblotting or protein blotting,

is an important technique in cell and molecular biology. In most basic terms, it is used to detect the presence of a specific protein in a complex mixture extracted from cells. The Western blotting procedure relies upon three key elements to accomplish this task: the separation of protein mixtures by size using gel electrophoresis; the efficient transfer of separated proteins to a solid support; and the specific detection of a target protein by appropriately matched antibodies. Once detected, the target protein will be visualized as a band on a blotting membrane, X-ray film, or an imaging system.

Since Western blotting is accomplished rapidly, using simple equipment

and inexpensive reagents, it is one of the most common laboratory

techniques. The results achieved are also easy to interpret, unique,

and unambiguous. Therefore, it is routinely used on its own, or along

with other immunoassays, in research and clinical settings.

The following technical bulletin is introduced for the entire western blotting procedure :
TECHNICAL BULLETIN EB07
THE COMPLETE WESTERN BLOTTING PROCEDURE
By: Joann Tye
(Modified from D. Douglas, S.P.J. Brooks, Pier Morin, and H. Mamady)
June 2005

The outlines of five days western blotting workshop are as follows:
1) Protein extraction
2) Determination of protein concentration
3) SDS-PAGE
4) Gel staining
5) Protein transfer
6) Immuno-detection of proteins
7) Tips for choosing antibody from companies
8) Quantification of protein bands by UV band software
9) Troubleshooting