Single-Channel Recording of Glycine Receptors in Human Embryonic Kidney (HEK) Cells

Andrew J R Plested; Jelena Baranovic

doi:10.1101/pdb.prot091652

Single-Channel Recording of Glycine Receptors in Human Embryonic Kidney (HEK) Cells

Research output: Contribution to journal › Article › peer-review

Abstract

This protocol describes how to record the single-channel activity of recombinant homomeric glycine receptors expressed in human embryonic kidney (HEK) cells. Cell-attached recordings readily reveal the large conductance (90 pS) and distinctive clusters of activations at high glycine concentration. This method for obtaining equilibrium recordings can be adapted to any ion channel receptor. The necessary extensions to outside-out patch for nonequilibrium recordings are also described, as are basic analyses of channel properties and activity.

Original language	English
Pages (from-to)	693-698
Number of pages	6
Journal	Cold Spring Harbor protocols
Volume	2016
Issue number	8
DOIs	https://doi.org/10.1101/pdb.prot091652
Publication status	Published - 1 Aug 2016

Keywords

Cells, Cultured
Epithelial Cells/physiology
Gene Expression
Humans
Kidney
Patch-Clamp Techniques/methods
Receptors, Glycine/analysis
Recombinant Proteins/analysis

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10.1101/pdb.prot091652

Jelena Baranovic
- School of Biological Sciences - Lecturer in Biochemistry
- SynthSys
Person: Academic: Research Active

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title = "Single-Channel Recording of Glycine Receptors in Human Embryonic Kidney (HEK) Cells",

abstract = "This protocol describes how to record the single-channel activity of recombinant homomeric glycine receptors expressed in human embryonic kidney (HEK) cells. Cell-attached recordings readily reveal the large conductance (90 pS) and distinctive clusters of activations at high glycine concentration. This method for obtaining equilibrium recordings can be adapted to any ion channel receptor. The necessary extensions to outside-out patch for nonequilibrium recordings are also described, as are basic analyses of channel properties and activity.",

keywords = "Cells, Cultured, Epithelial Cells/physiology, Gene Expression, Humans, Kidney, Patch-Clamp Techniques/methods, Receptors, Glycine/analysis, Recombinant Proteins/analysis",

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N2 - This protocol describes how to record the single-channel activity of recombinant homomeric glycine receptors expressed in human embryonic kidney (HEK) cells. Cell-attached recordings readily reveal the large conductance (90 pS) and distinctive clusters of activations at high glycine concentration. This method for obtaining equilibrium recordings can be adapted to any ion channel receptor. The necessary extensions to outside-out patch for nonequilibrium recordings are also described, as are basic analyses of channel properties and activity.

AB - This protocol describes how to record the single-channel activity of recombinant homomeric glycine receptors expressed in human embryonic kidney (HEK) cells. Cell-attached recordings readily reveal the large conductance (90 pS) and distinctive clusters of activations at high glycine concentration. This method for obtaining equilibrium recordings can be adapted to any ion channel receptor. The necessary extensions to outside-out patch for nonequilibrium recordings are also described, as are basic analyses of channel properties and activity.

KW - Cells, Cultured

KW - Epithelial Cells/physiology

KW - Gene Expression

KW - Humans

KW - Kidney

KW - Patch-Clamp Techniques/methods

KW - Receptors, Glycine/analysis

KW - Recombinant Proteins/analysis

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DO - 10.1101/pdb.prot091652

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JO - Cold Spring Harbor protocols

JF - Cold Spring Harbor protocols

SN - 1559-6095

IS - 8

ER -

Single-Channel Recording of Glycine Receptors in Human Embryonic Kidney (HEK) Cells

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Jelena Baranovic

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