Estrogen receptor protein, estrophilin, in a tissue sample is detected and quantified by radiochemical methods involving selective binding of the protein to controlled pore glass beads. Also disclosed are methods for selectively removing undesirable degradation products from radiochemically labelled non-polar steroid reagents, especially estradiol.
A hapten is obtained by replacing the 3-hydroxy group of equilin with HO--CO--A--O-- wherein A is an alkylene of one to six carbon atoms. The hapten is conjugated with an immunological carrier to provide an immunogen, which in turn produces a specific antiserum to equilin. The antiserum is used in a radioimmunoassay for equilin.
A hapten is obtained by replacing the 3-hydroxy group of 17 .alpha.-dihydroequilin with HO--CO--A--O-- wherein A is an alkylene of one to six carbon atoms. The hapten is conjugated with an immunological carrier to provide an immunogen, which in turn produces a specific antiserum to 17 .alpha.-dihydroequilin. The antiserum is used in a radioimmunoassay for 17 .alpha.-dihydroequilin.
The present invention provides unique prepared immunogens, site-specific polyclonal antisera and monoclonal antibodies against the DNA-binding domain of estrogen receptor protein, and immunoassay to determine the functional status of estrogen receptors in a cellular sample. Collectively or individually the component parts of the invention provide the ability not only to identify accurately the presence of human estrogen receptor but also the capability of determining whether the estrogen receptor exists in a functional or non-functional state.
An improved process of performing analysis methods based upon biospecific affinity reactions in heterogeneous systems, wherein a ligand dissolved in a liquid is reacted with a receptor immobilized on a carrier, characterized by utilizing as the carrier a porous matrix in the form of a material body having an open capillary pore system capable of absorbing and retaining liquid phase and having such limited void dimensions that the total reaction rate of the biospecific reaction or reactions within the pore system between ligand and immobilized receptor will be at least substantially independent of the diffusion of the ligand in the liquid phase. A porous matrix for use in said process has the characteristics mentioned above.
A process is disclosed for producing steroid hormone receptor samples to be utilized as controls during assays of various human tissue for steroid hormones, especially estrogen. The process comprises collecting tissue known to include such receptors, adding a buffer solution to the tissue, homogenizing the tissue and buffer solution, centrifuging the homogenized mixture, and thereafter collecting the supernatant. The supernatant which contains the desired receptors is subdivided into suitable control sample size and preferably lyophilized to a flake.
