Morris R J, Raisman G
J Neurochem. 1983 Mar;40(3):637-44. doi: 10.1111/j.1471-4159.1983.tb08028.x.
The conventional assay for measuring cell surface antigens--the quantitative absorption of antibody by tissue homogenates--proved inadequate when used to determine the level of Thy-1 glycoprotein in rat nerves and peripheral ganglia. In this paper we report that the binding of 125I-labelled Fab fragments of a monoclonal anti-Thy-1 antibody to cryostat sections is sufficiently sensitive to give consistent estimates of the Thy-1 level on single samples of even small nerves. Observed levels of Thy-1 were generally higher than had previously been thought, and in particular we found no nerves totally lacking the antigen. The lowest levels (6-10 pmol/mg protein) were in peripheral nerves with a large motor component. Autonomic and sensory nerves had higher levels (15-20 pmol/mg protein). The highest levels were on the optic nerve (34 pmol/mg protein), superior cervical sympathetic ganglion (40 pmol/mg protein), and the cerebellar vermis (46 pmoles/mg protein; the only brain region examined in this study). From a practical point of view, the cryostat assay has the advantage that measurements of Thy-1 can be done on sections from the same series as is used for immunohistochemical localisation.
用于测量细胞表面抗原的传统检测方法——通过组织匀浆对抗体进行定量吸收——在用于测定大鼠神经和外周神经节中Thy-1糖蛋白水平时被证明是不充分的。在本文中，我们报告了单克隆抗Thy-1抗体的125I标记Fab片段与低温恒温器切片的结合足够灵敏，能够对即使是小神经的单个样本中的Thy-1水平给出一致的估计。观察到的Thy-1水平通常比以前认为的要高，特别是我们发现没有完全缺乏该抗原的神经。最低水平（6 - 10 pmol/mg蛋白质）存在于具有大量运动成分的外周神经中。自主神经和感觉神经的水平较高（15 - 20 pmol/mg蛋白质）。最高水平存在于视神经（34 pmol/mg蛋白质）、颈上神经节（40 pmol/mg蛋白质）和小脑蚓部（46 pmoles/mg蛋白质；本研究中唯一检测的脑区）。从实际角度来看，低温恒温器检测法的优点是可以对与用于免疫组织化学定位相同系列的切片进行Thy-1的测量。
