Binding of estradiol-peroxidase conjugate to estrogen receptor

The binding of estradiol-horseradish peroxidase conjugate to rat uterine cytosolic estrogen receptor was studied. The conjugate having a steroid to enzyme ratio of 2.8:1 was allowed to bind to protamine precipitated receptor in presence or absence of 100-fold excess of free estradiol. The bound enzyme activity was measured and the data subjected to Scatchard analysis to obtain the dissociation constant and the number of binding sites. Although the binding parameter so obtained differed from values obtained using radiolabelled estradiol, the method may be used for comparative studies.