Estimation of Thy-1 in cryostat sections of nervous tissue.
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.
D000818:Animals; D000936:Antigen-Antibody Complex / Q000032:analysis; D000954:Antigens, Surface / Q000032:analysis*; D005260:Female; D007140:Immunoglobulin Fab Fragments / Q000276:immunology; D007158:Immunologic Techniques; D008722:Methods; D051379:Mice; D008808:Mice, Inbred CBA; D009420:Nervous System / Q000276:immunology*; D051381:Rats; D011919:Rats, Inbred Strains; D018800:Thy-1 Antigens
D000936:Antigen-Antibody Complex; D000954:Antigens, Surface; D007140:Immunoglobulin Fab Fragments; D018800:Thy-1 Antigens