Visualization of the multimeric structure of von Willebrand factor using a peroxidase-conjugated second antibody

We describe a technique for visualizing the multimeric structure of von Willebrand factor that does not require use of radioisotopes. This procedure uses a previously described discontinuous dodecyl sulfate agarose gel electrophoretic method followed by transfer of the protein to a nitrocellulose filter and detection by a double-antibody technique in which the second antibody is conjugated to horseradish peroxidase. In the presence of H2O2 and 4-chloro-1-napthol, the antigen appears as a series of about 15 colored bands. This method results in resolution of the high molecular weight components, does not require the use of radioisotopes, and can be completed in 2 days. It should be useful for diagnostic and investigational studies in clinical and research laboratories.