Utsunomiya Hirotoshi, Shan Liang, Kawano Ichiro, Iwasaki Arata, Ono Kazuo, Kobayashi Akira, Kuma Kanji, Kishikawa Seigo, Kakudo Kennichi
DVM, PhD.
Endocr Pathol. 1995 Autumn;6(3):223-227. doi: 10.1007/BF02739886.
The subcellular localization of parathyroid hormone (PTH) in the normal human parathyroid glands with particular reference to microwave antigen retrieval was investigated using peroxidase-labeled PTH antibody, immunohistochemical, and immunoelectron microscopic methods. The results revealed that PTH granules existed mainly as pro-PTH on the trans side of Golgi and in the regions adjacent to Golgi apparatus. Only a small proportion of secretory granules were stored near the plasma membrane. Microwave irradiation was essential for the immunodetection of PTH. As the irradiative time extended from 1 to 30 min, the staining intensity increased, and the subcellular preservation decreased. Microwave irradiation for 15 mm (with the sections in citrate buffer) with a power output of 500 W is the most ideal for PTH antigen retrieval, as well as for subcellular preservation.
利用过氧化物酶标记的甲状旁腺激素(PTH)抗体、免疫组织化学和免疫电子显微镜方法,研究了正常人甲状旁腺中PTH的亚细胞定位,特别参考了微波抗原修复。结果显示,PTH颗粒主要以前甲状旁腺激素原的形式存在于高尔基体反面膜囊和高尔基体附近区域。只有一小部分分泌颗粒储存在质膜附近。微波照射对于PTH的免疫检测至关重要。随着照射时间从1分钟延长至30分钟,染色强度增加,亚细胞保存效果下降。在柠檬酸盐缓冲液中对切片进行15分钟、功率输出为500瓦的微波照射,对于PTH抗原修复以及亚细胞保存来说是最理想的。