PURIFICATION OF ESTROGEN-RECEPTORS FROM MCF-7 HUMAN-BREAST CANCER-CELLS

  • 1 January 1980
    • journal article
    • research article
    • Vol. 40  (9) , 3172-3176
Abstract
Human estrogen receptor from MCF-7 breast cancer cells was partially purified to further characterize the chemical and biological properties of the receptor and to prepare specific receptor antibodies for radioimmunoassay. The estrogen receptor from MCF-7 cell cytosol was purified 1166-fold (27% recovery) over starting cytosol by combining (NH4)2SO4 sulfate precipitation, affinity chromatography and Sephadex G-100 gel filtration. The affinity resin consisted of estrone 17-(O-carboxymethyl)oxime:bovine serum albumin: Sepharose 4B. Under high salt conditions, the MW of the purified receptor is 50,000 and is identical to that obtained on chromatography of crude cytosol. Sucrose density gradient centrifugation also revealed the purified receptor sedimenting at the same position as the crude cytosol receptor. DNA-cellulose chromatography, hydroxylapatite chromatography, phosphocellulose chromatography, and diethylaminoethyl cellulose chromatography were much less effective purification methods.