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Updated: Jul 23, 2026

Detecting the Ligand-binding Domain Dimerization Activity of Estrogen Receptor Alpha Using the Mammalian Two-Hybrid Assay
Published on: December 19, 2018
The effect of dithiothreitol on the estrogen receptor
Abstract:
The effects of Dithiothreitol (DTT) on the estrogen receptor molecule were studied. DTT was evaluated for its potential abilities to preserve the integrity of the estrogen receptor. Storage at various extremes of pH was also considered for its possible adverse or positive effect. Cytosol was prepared utilizing rabbit uterus. Six aliquots of the cytosol were stored at 4 degrees C for 15 days. Two samples were adjusted to each pH, 6.0, 7.0, and 8.0, and to one sample of each pH was added DTT, 0.154 mg/ml. Aliquots of stored cytosols were analyzed for estrogen receptor (ER) on day 1, 3, 7, and 15 after adjusting the aliquots to pH 7.4. There was degradation of ER with time; degradation was less with DTT. The degradation also increases with the lower pH. The cytosol stored at 4 degrees C and pH 8.0 with DTT was the best environment in these experiments for stability of ER.
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