Estrogen receptor purification by affinity chromatography using an orange triazine dye.
A rapid two-step procedure was devised for the purification of the estrogen receptor from the calf uterus. A 900- to 1700-fold purification of the estrogen receptor was obtained using ammonium sulfate precipitation followed by dye affinity chromatography with Reactive Orange 14 immobilized to Sepharose. The Reactive Orange 14-Sepharose was used to purify the estrogen receptor in the presence or absence of estradiol as well as to purify the progesterone receptor. The purified estrogen receptor retained its estradiol- and DNA-binding properties and sedimented into sucrose gradients as the 5 S receptor dimer. The Reactive Orange 14-Sepharose is easily prepared and offers a higher yield and purity of the estrogen receptor than that afforded by estrogen- or heparin-Sepharose chromatography.
Bond
J P
JP
Department of Biophysics, University of Rochester School of Medicine and Dentistry, New York 14642.
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D000818:Animals; D002417:Cattle; D002846:Chromatography, Affinity; D003600:Cytosol / Q000032:analysis; D004247:DNA / Q000378:metabolism; D004958:Estradiol / Q000378:metabolism; D005260:Female; D066298:In Vitro Techniques; D011487:Protein Conformation; D011960:Receptors, Estrogen / Q000302:isolation & purification* / Q000378:metabolism; D012685:Sepharose; D014227:Triazines; D014599:Uterus / Q000032:analysis
D011960:Receptors, Estrogen; D014227:Triazines; C053575:reactive orange 14; D004958:Estradiol; D004247:DNA; D012685:Sepharose