Kendall M E, Hymer W C
Department of Molecular and Cell Biology, Pennsylvania State University, University Park 16802.
Endocrinology. 1987 Dec;121(6):2260-2. doi: 10.1210/endo-121-6-2260.
A new technique is described for quantification of PRL secretion from individual pituitary cells incubated 1-6 h on ImmobilonTM, a high protein binding capacity transfer membrane. After incubation, the membrane is processed according to procedures commonly used in Western blotting. Individual cells and their "zone of secretion" can be seen by light microscopy. Purified hormone, incubated and processed in a similar fashion, is used to generate a PRL standard curve. An OasysTM Image Analysis System is used to measure area and intensity of released hormone. PRL release ranged from 0.1-1.5 pg/cell, depending upon time of incubation and cell donor. Heterogeneity of hormone release from single cells was obvious.
本文描述了一种新技术，用于定量分析在ImmobilonTM（一种具有高蛋白结合能力的转移膜）上孵育1 - 6小时的单个垂体细胞分泌的催乳素（PRL）。孵育后，按照蛋白质印迹法常用的程序处理该膜。通过光学显微镜可以看到单个细胞及其“分泌区域”。以类似方式孵育和处理的纯化激素用于生成PRL标准曲线。使用OasysTM图像分析系统测量释放激素的面积和强度。PRL释放量在0.1 - 1.5 pg/细胞之间，具体取决于孵育时间和细胞供体。单个细胞激素释放的异质性很明显。
