What is western blotting?

1 Answer
Sep 5, 2016

It is the use of antibodies to detect proteins that have been separated by SDS-PAGE and then transferred to a membrane.


Cells contain a mixture of proteins (many thousands) and these can be separated by SDS-PAGE.

The proteins from the SDS-PAGE are then transferred, by the use of an electrical field, from the gel to a membrane typically made nitrocellulose or polyvinylidene fluoride (PVDF).

The membrane is probed with a primary antibody specific to the protein of interest. A secondary antibody is then used to detect the primary antibody. The secondary antibody will contain a marker that can be detected by a chemical reaction to either give a visible band on the membrane, or a signal that can be captured by a specialised camera.

If required I can expand on this answer in great depth as I have performed several hundred Western blots during my career.