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Elastin was isolated from the human aorta by three different extraction methods. Immunization was carried out in sheep. The presence of antibody against each elastin preparation in the sheep sera was confirmed by immunofluorescence. However, antiserum against elastin isolated by collagenase/guanidine dithioerythritol showed the most specific reactions in the cryostat sections. No cross-reactivity to type I, III and IV collagen, fibronectin, laminin and proteoglycan BM 1 was observed.

The antiserum against insoluble elastin from human aorta was applied to immuno-electron microscopy. In the preembedding method, only the outer surface of the amorphous component (elastin) of elastic fibers showed a positive reaction. A major problem encountered with the preembedding method is associated with the penetration of either the primary antiserum or the secondary antibody into the tissue and, in particular, into elastin. On the contrary, a positive reaction was observed inner zones of elastin in the postembedding method. While the postembedding method employed here has limitations with nonspecific binding to the embedding medium, the postembedding method offers a decided advantage over the preembedding method.