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. 1978 Sep;75(9):4359–4363. doi: 10.1073/pnas.75.9.4359

Ultrastructural antibody localization of alpha2-macroglobulin in membrane-limited vesicles in cultured cells.

M C Willingham, S S Yamada, I Pastan
PMCID: PMC336114  PMID: 81488

Abstract

We have been developing a procedure for localizing intracellular antigens in cultured cells, by using peroxidase-labeled antibodies, that allows good morphologic preservation. Although useful, our previous technique did not preserve the morphology of membranes, and the location of the peroxidase reaction product was difficult to establish. In this paper, we report major improvements on the basic technique that markedly enhance the quality of localization and of morphology. Saponin is used to permeabilize membranes without destroying their morphology. The amount of reaction product is enhanced with a peroxidase-antiperoxidase label. The clarity of morphologic detail and contrast of reaction product density are increased by using postsectioning staining with the osmium/thiocarbohydrazide/osmium and uranyl acetate/lead citrate procedures. We have applied this technique to the ultrastructural localization of alpha2-macroglobulin and demonstrated that it is localized in membrane-limited vesicles. We have also used this method to improve the preservation of structures for localization by fluorescence microscopy.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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