An assay procedure for solubilized thyroid hormone receptor: use of Lipidex.
A method using Lipidex-1000 is reported for the assay of thyroid hormone receptor activity. The receptor was extracted from rat liver nuclei, incubated with [125I]T3, and applied to 1-ml disposable pipet tip columns containing a small volume of Lipidex. The resin absorbed the free hormone, and the receptor-bound hormone was recovered in the eluate. The method allowed accurate assay of the receptor activity, satisfying a linear relationship between the activity and the receptor protein concentration. The usefulness of this method was demonstrated by determining the pH optimum and the Kd and Bmax of T3 binding by the receptor. The Lipidex column can be used for the preparation of an unoccupied receptor after dissociation of the endogenously bound hormone.
Li
Q
Q
Department of Biochemistry, Osaka City University Medical School, Japan.
Yamamoto
N
N
Inoue
A
A
Morisawa
S
S
D000042:Absorption; D000818:Animals; D001665:Binding Sites; D006863:Hydrogen-Ion Concentration; D007457:Iodine Radioisotopes; D008099:Liver / Q000737:chemistry; D008851:Micropore Filters; D051381:Rats; D011988:Receptors, Thyroid Hormone / Q000187:drug effects / Q000378:metabolism*; D012116:Resins, Plant; D012965:Sodium Chloride / Q000494:pharmacology; D012995:Solubility; D014284:Triiodothyronine / Q000378:metabolism
D007457:Iodine Radioisotopes; D011988:Receptors, Thyroid Hormone; D012116:Resins, Plant; D014284:Triiodothyronine; D012965:Sodium Chloride