Abstract
A quantitative immunoprecipitation assay was conducted to study the developmental expression of nerve growth factor (NGF) receptor by using a monoclonal antibody (192-IgG) against rat NGF receptor. Either homogenates or plasma membrane enriched preparations from isolated tissues or whole rat embryos were bound with 125I-NGF and crosslinked by 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide. The receptor was then solubilized by 2% octylglucoside, bound with 192-IgG, and precipitated by secondary antibodies against mouse IgG prebound to Pansorbin. The complex, 125I-NGF/receptor/antibodies/Pansorbin, was separated from free ligand by rapid centrifugation through a 0.15 M sucrose cushion. The tube was frozen, and the tip containing the radioactivity was cut and counted. In some cases, the pellet was resuspended in SDS-PAGE sample buffer and subjected to a SDS-PAGE autoradiographic analysis.
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© 1988 Kluwer Academic Publishers
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Yan, Q., Johnson, E.M. (1988). A Quantitive Study of the Developmental Expression of Nerve Growth Factor Receptors in Rats. In: Ferrendelli, J.A., Collins, R.C., Johnson, E.M. (eds) Neurobiology of Amino Acids, Peptides and Trophic Factors. Topics in the Neurosciences, vol 8. Springer, Boston, MA. https://doi.org/10.1007/978-1-4613-1721-0_27
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DOI: https://doi.org/10.1007/978-1-4613-1721-0_27
Publisher Name: Springer, Boston, MA
Print ISBN: 978-1-4612-8969-2
Online ISBN: 978-1-4613-1721-0
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