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An analysis of the decrease in the assayed level of charged bovine estrogen receptor observed at physiological ionic

F V Peale, J Bond, R Hilf

    Journal of Steroid Biochemistry
    |February 1, 1985
    PubMed
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    Heifer uterine estrogen receptor (ER) levels are underestimated at moderate ionic strength due to resin adsorption. Trypsin or molybdate treatment prevents this ER assay reduction.

    Area of Science:

    • Reproductive biology
    • Endocrinology
    • Biochemistry

    Background:

    • Estrogen receptor (ER) levels are crucial for reproductive health.
    • Accurate ER quantification is essential for understanding uterine function.

    Purpose of the Study:

    • To investigate the cause of reduced estrogen receptor (ER) levels measured at physiological ionic strength.
    • To identify factors affecting ER binding to assay resins.

    Main Methods:

    • Assaying heifer uterine ER levels using [3H]estradiol with Dextran-coated charcoal or gel filtration.
    • Varying ionic strength (0-50 mM, 150-200 mM, 400-450 mM) during ER separation.
    • Treating ER with trypsin or molybdate before assay.

    Main Results:

    Related Experiment Videos

    • ER levels were approximately halved at 150-200 mM ionic strength compared to lower or higher ionic strengths.
    • Treatment with trypsin or molybdate prevented the reduction in assayed ER levels.
    • Evidence suggests preferential adsorption of ER to assay resins at moderate ionic strength.

    Conclusions:

    • The observed decrease in ER levels at physiological ionic strength is due to preferential adsorption to assay resins.
    • A hydrophobic region on the ER, susceptible to trypsin cleavage, mediates this adsorption.
    • These findings highlight the importance of ionic strength control in ER assays.