The use of immobilized estradiol antiserum in the study of receptors and other estradiol-binding proteins

✍ Scribed by Jerry H. Fishman; Jack Fishman

• Publisher: Elsevier Science
• Year: 1979
• Tongue: English
• Weight: 691 KB
• Volume: 94
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(79)90361-0

✦ Synopsis

Antiestradiol

antibody immobilized on a polymer film is set in competition for tritium-labeled estradiol with estradiol receptor or antiestradiol antibody in solution. Measurement of equilibrium quantity of radiolabel in test solutions in the presence and in the absence of dissolved antibody or receptor gives a data base for evaluating the association constant and the quantity of the estradiol-complexing species in solution. The immobilized antibody method avoids the problems and uncertainties arising in the step of separating the bound from the free hormone which is the pivotal step in all currently used procedures for the determination and characterization of steroid receptors. With the exception of equilibrium dialysis, it is the only procedure presently available to study steroid antibodies and receptors in solution at unperturbed equilibrium. Using the immobilized antibody method at 4"C, 0.86 ? 0.12 x 10" M-' was the association constant found for estradiol receptor in rat uterine cytosol, and 2.6 ? 0.5 x 10" M-' was the association constant found for antiestradiol antibody raised in a rabbit to estradiol linked to bovine serum albumin via a C-6 carboxymethyloxime.