Verifying Transfected Protein Expression by Western Blotting

Western blotting involves separating the proteins in a cell extract on the basis of size on a polyacrylamide gel (17), transferring these proteins onto a blotting membrane (18,19), and then probing this blot using an antibody specific to the transfected protein (21). Western blotting provides a means of estimating the expression level of the transfected protein, and whether the expressed protein is of the expected size. It is most useful in such cases where, for example, a functional wild-type protein is compared with a nonfunctional mutant in a reporter assay, to verify that the functional wild-type and nonfunctional mutant proteins are expressed at similar levels in the respective transfected cells.

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