Homogeneous bivalent insulin receptor: Purification using insulin coupled to 1,1′-carbonyldiimidazole activated-agarose

A novel affinity gel, consisting of insulin coupled to 1,1′-carbonyldiimidazole-activated agarose (CDI-agarose), was used to purify insulin receptors from human placenta to homogeneity. This affinity gel is reproducibly prepared and is reported to have a number of advantages over the standard cyanog...

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Veröffentlicht in:	Biochemical and biophysical research communications 1985-11, Vol.132 (3), p.1059-1065
Hauptverfasser:	Newman, Julie D., Harrison, Leonard C.
Format:	Artikel
Sprache:	eng
Schlagworte:	Chromatography, Affinity Electrophoresis, Polyacrylamide Gel Female Humans Imidazoles - pharmacology insulin Iodine Radioisotopes man Molecular Weight placenta Placenta - analysis Pregnancy Receptor, Insulin - isolation & purification Sepharose
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container_title	Biochemical and biophysical research communications
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creator	Newman, Julie D. Harrison, Leonard C.
description	A novel affinity gel, consisting of insulin coupled to 1,1′-carbonyldiimidazole-activated agarose (CDI-agarose), was used to purify insulin receptors from human placenta to homogeneity. This affinity gel is reproducibly prepared and is reported to have a number of advantages over the standard cyanogen bromide activated supports, such as ease and simplicity of coupling and minimal ligand leakage and non-specific binding. Insulin receptors in Triton X-100-solubilized microsomal membranes were purified 2,000-fold by sequential affinity chromatography on wheat germ lectin-agarose and insulin-CDI-activated agarose. They have one of the highest specific insulin-binding capacities (6 nmol/mg protein) reported and can be calculated to have a binding valence of two on the basis that the molecular weight of the oligomeric receptor is 300–350,000.
doi_str_mv	10.1016/0006-291X(85)91914-X
format	Article
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source	MEDLINE; Elsevier ScienceDirect Journals
subjects	Chromatography, Affinity Electrophoresis, Polyacrylamide Gel Female Humans Imidazoles - pharmacology insulin Iodine Radioisotopes man Molecular Weight placenta Placenta - analysis Pregnancy Receptor, Insulin - isolation & purification Sepharose
title	Homogeneous bivalent insulin receptor: Purification using insulin coupled to 1,1′-carbonyldiimidazole activated-agarose
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