Purification of estrogen receptors from MCF-7 human breast cancer cells

M. T. Chong, Marc E Lippman

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

We have partially purified human estrogen receptor from MCF-7 breast cancer cells in order 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 1,166-fold (27% recovery) over starting cytosol by combining ammonium 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 molecular weight 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. The following methods to purify crude cytosol estrogen receptors are much less effective. (a) With DNA-cellulose chromatography, KCl elution revealed peaks at 0.04 m KCl (11-fold purification) and 0.22 m KCl (12.5-fold purification). This could not be combined sequentially with affinity chromatography because high salt conditions were required for affinity elution, and removal of salt by dialysis caused the receptor to adhere to the tubing. (b) With hydroxylapatite chromatography, phosphate elution revealed peaks at 0.12 m phosphate (1.1-fold purification) and 0.175 m phosphate (1.1-fold purification). (c) With phosphocellulose chromatography, KCl elution revealed only one peak at 0.18 m KCl (1.6-fold purification). (d) With diethylaminoethyl cellulose chromatography, KCl elution revealed 4 peaks at 0.025 m (1-fold purification), 0.12 m (1.27-fold purification), 0.14 m (1.28-fold purification), and 0.2 m (0.36-fold purification).

Original languageEnglish
Pages (from-to)3172-3176
Number of pages5
JournalCancer Research
Volume40
Issue number9
StatePublished - Jan 1 1980
Externally publishedYes

Fingerprint

Estrogen Receptors
Cytosol
Chromatography
Breast Neoplasms
Salts
Phosphates
Affinity Chromatography
Oximes
Density Gradient Centrifugation
Estrone
MCF-7 Cells
Ammonium Sulfate
Durapatite
Bovine Serum Albumin
Cellulose
Sepharose
Radioimmunoassay
Gel Chromatography
Sucrose
Dialysis

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Chong, M. T., & Lippman, M. E. (1980). Purification of estrogen receptors from MCF-7 human breast cancer cells. Cancer Research, 40(9), 3172-3176.

Purification of estrogen receptors from MCF-7 human breast cancer cells. / Chong, M. T.; Lippman, Marc E.

In: Cancer Research, Vol. 40, No. 9, 01.01.1980, p. 3172-3176.

Research output: Contribution to journalArticle

Chong, MT & Lippman, ME 1980, 'Purification of estrogen receptors from MCF-7 human breast cancer cells', Cancer Research, vol. 40, no. 9, pp. 3172-3176.
Chong, M. T. ; Lippman, Marc E. / Purification of estrogen receptors from MCF-7 human breast cancer cells. In: Cancer Research. 1980 ; Vol. 40, No. 9. pp. 3172-3176.
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