Protein purification using a soluble affinity matrix: purification of estrogen receptor with estradiol-polylysine conjugate.
A new strategy for protein purification using a soluble affinity matrix is described. The method was used for purification of estrogen receptor. Cytosols from rat uteri and human fibroid uterine tissue, after fractionation by ammonium sulfate, were treated with estradiol-polylysine conjugate. The highly basic affinity complex was separated from other proteins by DEAE-Sephacel chromatography. After dissociation of the eluted complex with excess estradiol, the receptor was recovered by CM-Sephadex chromatography. A 2000-fold purification of the rat uterine estrogen receptor was obtained with an activity recovery of 35%.
Bhattacharjee
M
M
Indian Institute of Chemical Biology, Calcutta.
Ali
E
E
D000818:Animals; D002458:Cell Fractionation; D002846:Chromatography, Affinity / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D004958:Estradiol / Q000378:metabolism*; D005260:Female; D011107:Polylysine / Q000378:metabolism*; D051381:Rats; D011919:Rats, Inbred Strains; D011960:Receptors, Estrogen / Q000302:isolation & purification* / Q000378:metabolism; D012995:Solubility
D011960:Receptors, Estrogen; D011107:Polylysine; D004958:Estradiol