avidin biotinylated horseradish-peroxidase complex method

avidin biotinylated horseradish-peroxidase complex method

A commercial method for detecting antigen or antibody in tissues involving avidin, a 68-kD egg-white glycoprotein with a very high natural affinity and multiple binding sites for biotin, a vitamin readily bound covalently to an antibody; the ABC system allows amplification of antigen “signal” obtained by the multiple binding sites available on the avidin; the avidin:biotin linkage can be used for gene mapping, double label studies, DNA in situ hybridisation, hybridoma screening, Southern blotting, RIA, solid-phase ELISA, immune electron microscopy, studies of neuronal transport, and as a means of controlling enzymatic reactions.
 
Method
The tissue containing the antigen of interest is frozen, air-dried and acetone fixed, incubated with a primary antibody, washed, then incubated with a secondary antibody bearing attached biotin, washed, re-incubated with fluorescently or enzyme-tagged avidin, counterstained, and examined by light microscopy.

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