Transfer Buffers & Detection Reagents

Blotting of nucleic acids is usually performed as a capillary blot using SSC or SSPE as blotting buffer.
The proteins studied by Western blotting are most often separated by acrylamide gel electrophoresis. As these gels are too restrictive for capillary blotting, Western blots are most often done by electrotransfer. Western blotting usually involves considerable handling of the gels and a support matrix of some sort is a great help especially with low concentration acrylamide gels. Most protein transfer is done from SDS-PAGE gels. It is also possible to transfer from IEF gels. Depending on the type of blotting (tank or semi-dry blotting) and the specific characteristics of the proteins different buffers may be used to Western Blotting. For small proteins or for tank blotting after SDS PAGE the Towbin buffer is recommended, for blotting proteins after SDS PAGE or IEF by semi-dry blotting, a discontinuous buffer system, is recommended.