General immunostain of blotted proteins

Wojtkowiak et al. (1983) devised a simple, universal and very sensitive, but time-consuming method for the detection of proteins on blots. 

After blotting of the proteins, they are dinitrophenylated (Section 12.5). This hapten is then detected with anti-DNP antibodies (Miles, Inc.j by one of the methods discussed below. Detectability by this method is at least equal to that of autoradiography ( 1 ng). The intensity of the staining, however, is not representative of the stoichiometry of the protein in the blot. Kittler et al. (1984) haptenated the amino groups of the blotted proteins with pyridoxal-5 -phosphate. This method requires no organic solvents and can also be used with Zetabind membranes. 

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