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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.
Transfer Buffers & Detection Reagents
