Press M F, Greene G L
Lab Invest. 1984 Apr;50(4):480-6.
We have used monoclonal antiestrophilin antibodies to develop an improved immunocytochemical method for localizing estrogen receptors in tissue sections with an indirect immunoperoxidase technique. Rat monoclonal antibodies were raised against human estrogen receptor (estrophilin) protein derived from the cytosol of MCF-7 human breast cancer cells. These monoclonal antibodies have been shown to have extensive cross-reactivity with estrogen receptors from various primate and nonprimate tissues. We have used frozen sections of human proliferative phase endometrium and an indirect immunoperoxidase technique to establish conditions for demonstrating estrogen receptor antigenic determinants in frozen tissue sections. The method involves (a) brief fixation with formaldehyde-containing fixatives either prior to freezing or immediately after cutting cryostat sections, (b) bleaching the tissue of endogenous peroxidase activity, (c) application of primary antibody or control immunoglobulin, (d) application of an adsorbed bridging antibody (goat antirat IgG), and (e) application of rat peroxidase-antiperoxidase followed by diaminobenzidine. Specific nuclear staining for estrogen receptor antigenic determinants was observed in the vast majority of epithelial and stromal cells. No specific cytoplasmic staining was identified in cryostat sections of any of the 17 cases studied for this report.
我们已使用抗雌激素受体单克隆抗体，通过间接免疫过氧化物酶技术开发出一种改进的免疫细胞化学方法，用于在组织切片中定位雌激素受体。针对源自MCF-7人乳腺癌细胞胞质溶胶的人雌激素受体（雌激素受体蛋白）制备了大鼠单克隆抗体。这些单克隆抗体已被证明与来自各种灵长类和非灵长类组织的雌激素受体具有广泛的交叉反应性。我们使用人增殖期子宫内膜的冷冻切片和间接免疫过氧化物酶技术，来确定在冷冻组织切片中显示雌激素受体抗原决定簇的条件。该方法包括：（a）在冷冻前或用低温恒温器切片后立即用含甲醛的固定剂进行短暂固定；（b）去除组织的内源性过氧化物酶活性；（c）应用一抗或对照免疫球蛋白；（d）应用吸附的桥连抗体（山羊抗大鼠IgG）；（e）应用大鼠过氧化物酶-抗过氧化物酶，然后用二氨基联苯胺。在绝大多数上皮细胞和基质细胞中观察到雌激素受体抗原决定簇的特异性核染色。在本报告研究的17例病例的任何低温恒温器切片中均未发现特异性细胞质染色。
