Brown W J, Farquhar M G
Section of Biochemistry, Molecular, and Cell Biology, Cornell University, Ithaca, New York 14853.
Methods Cell Biol. 1989;31:553-69. doi: 10.1016/s0091-679x(08)61626-x.
We have presented our detailed methods for localizing antigens in cultured cells and tissue sections by IP at the EM level. Immunoperoxidase cytochemistry is particularly well suited for the study of sparse antigens as a result of the enzymatic amplification afforded by the method, and of molecules confined within a membrane-enclosed compartment wherein the DAB reaction produce can accumulate. Although IP is commonly used to localize membrane-compartmentalized molecules, reliable qualitative information can also be obtained on cytoplasmic antigens as well (Anderson et al., 1978; Merisko et al., 1986; Rodman et al., 1984). For these and other reasons, it is likely that IP cytochemistry will continue to be an important tool for the cell biologist especially in the study of membrane traffic. Other inventive combinations of immunocytochemical methods will likely be forthcoming, for example, combining IP localization with postembedding labeling by colloidal-gold conjugates to provide triple EM labeling.
我们已经展示了在电子显微镜水平下通过免疫过氧化物酶法在培养细胞和组织切片中定位抗原的详细方法。免疫过氧化物酶细胞化学特别适合研究稀疏抗原，这是由于该方法提供的酶促放大作用，以及对于局限于膜封闭隔室内的分子，其中二氨基联苯胺（DAB）反应产物可以积累。尽管免疫过氧化物酶法通常用于定位膜分隔的分子，但也可以获得关于细胞质抗原的可靠定性信息（安德森等人，1978年；梅里斯科等人，1986年；罗德曼等人，1984年）。由于这些以及其他原因，免疫过氧化物酶细胞化学可能仍然是细胞生物学家的重要工具，特别是在膜运输研究中。免疫细胞化学方法的其他创新组合可能即将出现，例如，将免疫过氧化物酶法定位与胶体金缀合物的包埋后标记相结合，以提供三重电子显微镜标记。
