Cell membrane coating with glutaraldehyde: Application to a versatile solid-phase assay for thyroid membrane proteins and molecules interacting with thyroid membranes

In defined conditions, glutaraldehyde was shown to tightly bind cell membranes to flexible microtiter plates without significant alteration of the antigenic and functional properties of membrane proteins. In the presence of 0.06% glutaraldehyde, human thyroid membranes were bound to plastic firmly e...

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Veröffentlicht in:Analytical biochemistry 1985-08, Vol.148 (2), p.320-327
Hauptverfasser: Adler, Grazyna, Ruf, Jean, Benkirane, Moufid, Ferrand, Mireille, Carayon, Pierre
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container_end_page 327
container_issue 2
container_start_page 320
container_title Analytical biochemistry
container_volume 148
creator Adler, Grazyna
Ruf, Jean
Benkirane, Moufid
Ferrand, Mireille
Carayon, Pierre
description In defined conditions, glutaraldehyde was shown to tightly bind cell membranes to flexible microtiter plates without significant alteration of the antigenic and functional properties of membrane proteins. In the presence of 0.06% glutaraldehyde, human thyroid membranes were bound to plastic firmly enough to resist numerous washing and flicking steps; the coated membranes remained almost unaltered with regard to monoclonal antibody and thyrotropin binding as well as adenylate cyclase and peroxidase activities. Based on the use of thyroid membrane-coated microtiter plates, a versatile solid-phase assay was developed which allowed screening of anti-membrane monoclonal antibodies, detection of thyrotropin-displacing activity in hormone and antibody preparations, and monitoring of fractionation experiments of solubilized membrane antigens and thyrotropin receptor. It was concluded that the use of glutaraldehyde for coating cell membranes to flexible microtiter plates enabled the establishment of simple, rapid, and reliable assays for detection and quantitation of membrane proteins and molecules interacting with membranes.
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subjects Adenylyl Cyclases - metabolism
Aldehydes
antibodies
Antibodies, Monoclonal
Antigens - analysis
Applied sciences
binding assays
Biological and medical sciences
Cell Membrane - metabolism
Cell membranes. Ionic channels. Membrane pores
Cell structures and functions
Exact sciences and technology
Fundamental and applied biological sciences. Psychology
Glutaral
glutaraldehyde
hormones
Humans
Iodide Peroxidase - metabolism
man
membrane proteins
Membrane Proteins - analysis
Membrane Proteins - immunology
Membrane Proteins - metabolism
Microchemistry
Molecular and cellular biology
Other techniques and industries
receptors
Receptors, Cell Surface - metabolism
Receptors, Thyrotropin
Solubility
thyroid
Thyroid Gland - metabolism
title Cell membrane coating with glutaraldehyde: Application to a versatile solid-phase assay for thyroid membrane proteins and molecules interacting with thyroid membranes
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