Abstract
The detection of expression of oncogene products in human neoplasms requires fresh tissue for karyotyping, DNA and RNA extraction and immunoblotting. Immunostaining of formalin-fixed paraffin embedded sections provides a reliable and more feasible alternative, which permits study of archival material. Twenty high grade astrocytomas and twenty-nine low grade astrocytomas were examined by immunostaining for expression of EGF-R, c-myc, c-myb, N-ras/c-Ha-ras and c-fos oncoproteins. A comparison of the relative efficacy of peroxidase - antiperoxidase (PAP) and colloidal gold immunostaining (IG) revealed greater sensitivity of IG at antibody dilutions of 1:500 and 1:600. The detection of antigens at lower levels of expression is possible with IG because of one-to-one binding of colloidal gold markers to antigens. Additional advantages of IG over PAP immunohistochemistry include potential semiquantitative applications to electron microscopy and subcellular localization of antigens. A modified immunogold-silver technique was developed for application to the nervous system.
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