Cell-based enzyme-linked immunosorbent assay (cell-ELISA) analysis of native and recombinant glutamate receptors

Elek Molnár

doi:10.1007/978-1-4939-9077-1_4

Cell-based enzyme-linked immunosorbent assay (cell-ELISA) analysis of native and recombinant glutamate receptors

Elek Molnár^*

^*Corresponding author for this work

Research output: Chapter in Book/Report/Conference proceeding › Chapter in a book

1 Citation (Scopus)

Abstract

Glutamate receptors (GluRs) located primarily on the membranes of neurons and glial cells are responsible for excitatory synaptic transmission in the central nervous system. The transport of GluRs to the cell surface is a highly regulated dynamic process that determines neuronal excitability and synaptic responses. The molecular and cellular mechanisms of GluR trafficking are often studied in cell cultures. These studies require sensitive techniques that allow the measurement of total and surface expressed GluRs in cell populations. The cell-based enzyme-linked immunosorbent assay (cell-ELISA) combines steps of direct immunochemical labelling of cell cultures and ELISA. It can be used for quantitative comparisons of surface expressed and total protein contents of various cell cultures. While several cell-ELISA protocols are available for different cell types, in this chapter we describe the procedure that we have applied for the investigation of quantitative changes in the cell surface expression of recombinant ionotropic glutamate receptors (iGluRs) in adherent human embryonic kidney 293 (HEK293) cells and endogenous iGluR proteins in primary neuronal cultures.

Original language	English
Title of host publication	Methods in Molecular Biology
Subtitle of host publication	Glutamate receptors: Methods and protocols
Publisher	Springer
Chapter	4
Pages	47-54
Number of pages	8
ISBN (Print)	978-1-4939-9077-1
DOIs	https://doi.org/10.1007/978-1-4939-9077-1_4 https://doi.org/10.1007/978-1-4939-9077-1_4
Publication status	Published - 23 Mar 2019

Publication series

Name	Methods in Molecular Biology
Volume	1941
ISSN (Print)	1064-3745

Keywords

Antibodies
Cell-ELISA
Cell surface protein
Glutamate receptor
Neuronal culture
Targeting
trafficking
Neurochemistry
Receptors
Membrane proteins
Cell biology
Cell culture

Access to Document

10.1007/978-1-4939-9077-1_4Licence: Unspecified
10.1007/978-1-4939-9077-1_4

Link to publication in Scopus

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Projects

1 Finished

Development of biotin-tagged affinity ligands and fluorophore-conjugated probes for the study of native kainate receptors

Molnar, E.

1/03/13 → 1/03/16

Project: Research

Cite this

Molnár, E. (2019). Cell-based enzyme-linked immunosorbent assay (cell-ELISA) analysis of native and recombinant glutamate receptors. In Methods in Molecular Biology: Glutamate receptors: Methods and protocols (pp. 47-54). (Methods in Molecular Biology; Vol. 1941). Springer. https://doi.org/10.1007/978-1-4939-9077-1_4, https://doi.org/10.1007/978-1-4939-9077-1_4

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abstract = "Glutamate receptors (GluRs) located primarily on the membranes of neurons and glial cells are responsible for excitatory synaptic transmission in the central nervous system. The transport of GluRs to the cell surface is a highly regulated dynamic process that determines neuronal excitability and synaptic responses. The molecular and cellular mechanisms of GluR trafficking are often studied in cell cultures. These studies require sensitive techniques that allow the measurement of total and surface expressed GluRs in cell populations. The cell-based enzyme-linked immunosorbent assay (cell-ELISA) combines steps of direct immunochemical labelling of cell cultures and ELISA. It can be used for quantitative comparisons of surface expressed and total protein contents of various cell cultures. While several cell-ELISA protocols are available for different cell types, in this chapter we describe the procedure that we have applied for the investigation of quantitative changes in the cell surface expression of recombinant ionotropic glutamate receptors (iGluRs) in adherent human embryonic kidney 293 (HEK293) cells and endogenous iGluR proteins in primary neuronal cultures.",

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KW - Cell surface protein

KW - Glutamate receptor

KW - Neuronal culture

KW - Targeting

KW - trafficking

KW - Neurochemistry

KW - Receptors

KW - Membrane proteins

KW - Cell biology

KW - Cell culture

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