Detection of cell surface dopamine receptors

Jiping Xiao, Clare Bergson

Research output: Chapter in Book/Report/Conference proceedingChapter

1 Scopus citations

Abstract

Dopamine receptors are a class of metabotropic G protein-coupled receptors. Plasma membrane expression is a key determinant of receptor signaling, and one that is regulated both by extra and intracellular cues. Abnormal dopamine receptor signaling is implicated in several neuropsychiatric disorders, including schizophrenia and attention deficit hyperactivity disorder, as well as drug abuse. Here, we describe in detail the application of two complementary applications of protein biotinylation and enzyme-linked immunoabsorbent assay (ELISA) for detecting and quantifying levels of dopamine receptors expressed on the cell surface. In the biotinylation method, cell surface receptors are labeled with Sulfo-NHS-biotin. The charge on the sulfonyl facilitates water solubility of the reactive biotin compound and prevents its diffusion across the plasma membrane. In the ELISA method, surface labeling is achieved with antibodies specific to extracellular epitopes on the receptors, and by fixing the cells without detergent such that the plasma membrane remains intact.

Original languageEnglish (US)
Title of host publicationDopamine
Subtitle of host publicationMethods and Protocols
PublisherHumana Press Inc.
Pages3-13
Number of pages11
ISBN (Print)9781627032506
DOIs
StatePublished - 2013

Publication series

NameMethods in Molecular Biology
Volume964
ISSN (Print)1064-3745

Keywords

  • ADHD
  • Biotinylation
  • DAPI
  • ELISA
  • Plasma membrane
  • Schizophrenia

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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  • Cite this

    Xiao, J., & Bergson, C. (2013). Detection of cell surface dopamine receptors. In Dopamine: Methods and Protocols (pp. 3-13). (Methods in Molecular Biology; Vol. 964). Humana Press Inc.. https://doi.org/10.1007/978-1-62703-251-3_1