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Measurement of human nerve growth factor binding sites in brain and in peripheral tissues by a specific immunoprecipitation assay

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Summary

Using a monoclonal antibody against the human nerve growth factor (NGF) receptor (20.4 IgG), a specific immunoprecipitation assay for the quantitation of human NGF binding sites has been established. The procedure involves specific labeling of NGF receptors by covalent crosslinking to 125I-NGF with a carbodiimide reagent, and subsequent immunoprecipitation of the detergent-extracted 125I-NGF-receptor complexes with the 20.4 IgG. This two-site assay provides a specific method to measure NGF binding sites in peripheral tissues and central nervous system. Moreover, it allows analysis of the immunoprecipitated receptor species by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The finding that specific NGF binding sites are expressed on human neuronal and nonneuronal tissues, including lymphoid tissues, indicates that NGF exerts a broader physiological function than originally ascribed.

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Abbreviations

BSA:

bovine serum albumin

NGF:

nerve growth factor

PAGE:

polyacrylamide gel electrophoresis

PBS:

phosphatebuffered saline

SCG:

superior cervical ganglion

SDS:

sodium dodecyl sulfate

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Authors and Affiliations

  1. Deptartment of Physiology, University of Basel, Vesalianum, Vesalgasse 1, CH-4051, Basel, Switzerland

    P. Ehrhard & U. Otten

  2. Institute of Pathology, University of Basel, Schönbeinstrasse 40, CH-4003, Basel, Switzerland

    F. Gudat

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  1. P. Ehrhard
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  2. F. Gudat
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  3. U. Otten
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Ehrhard, P., Gudat, F. & Otten, U. Measurement of human nerve growth factor binding sites in brain and in peripheral tissues by a specific immunoprecipitation assay. Naunyn-Schmiedeberg's Arch Pharmacol 342, 251–257 (1990). https://doi.org/10.1007/BF00169434

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  • DOI: https://doi.org/10.1007/BF00169434

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