= Western blot antibody detection
Blocking occupies unused binding sites on the <Western blot membrane>. The <primary antibody> is applied, unbound material is washed away, and a labelled <secondary antibody> detects retained <primary antibody>. A <fluorescence> label emits detectable <light> after excitation; an <enzyme>-linked label can instead generate a colored product or <light>. Washing reduces background but does not prove that remaining binding is selective.
In the original experiment, blocking used excess <protein>. An anti-<ribosome>-<protein> serum followed by labelled <secondary antibody> produced colored or <fluorescence> bands; preimmune serum supplied comparison strips. The color readout was an <enzyme> reaction product, rather than a measurement of target <enzyme> activity. <Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications by Harry Towbin and others>[Towbin, Staehelin and Gordon, pp. 4351–4353: Immunological Detection of Proteins on Nitrocellulose and Figure 4, including preimmune-serum controls]{ref}
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