Protein Blotting: Principles and Applications
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Protein blotting was initially introduced in the late 1970s to identify protein antigens that bound to spectfic antibodies
(1
,2
) In this procedure, a complex protein fraction was separated by electrophoresis and the proteins transferred and bound to
a membrane, which was then probed with a radiolabeled antibody This concept and procedure were based on blotting and hybridization
methods developed for DNA by Southern (3
) (and logically called Southern blotting), which was subsequently modified for RNA (4
) (less logtcally called Northern blotting by some authors). It is, therefore, not surprising that the protem/antibody procedure
was called Western blotting (2
), and that Southwestern (protein/DNA) and Far Western (or West Western) (protem/protem) procedures have since been developed
In addition, it is possible to apply proteins to the membrane in solution (dot-blotting), or to transfer them directly from
tissues (squash blots and tissue prints), whereas the development of microscale protein sequencing and ammo-acid analysts
has allowed the use of blotting to purify and characterize individual components of highly complex mixtures