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Estimation of Thy‐1 in Cryostat Sections of Nervous Tissue
Author(s) -
Morris R. J.,
Raisman G.
Publication year - 1983
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1111/j.1471-4159.1983.tb08028.x
Subject(s) - cryostat , nervous system , nervous tissue , anatomy , biology , neuroscience , physics , superconductivity , quantum mechanics
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 125 1‐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.