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US 4,375,459 AGranted
Cytoreceptor assay
AssigneeThe Regents of the University of California
InventorStavros C. Manolagas
Life of this patent
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Abstract
Novel assays are provided employing specific ells having cytoreceptors for an analyte. The assay combines the specific cells, the sample, and a tagged analyte, normally radioactively tagged. After a sufficient time, the cells are separated, conveniently by centrifugation, and either the supernatant or the cells assayed for the signal resulting from the tag.
Claims14
A method for determining an analyte capable of specifically binding to a cytosol cell receptor, said method comprising:
- combining in a liquid assay medium supportive of cell viability, a sample suspected of containing said analyte, cells having a specific extracellular receptor for said analyte, and tagged analyte, wherein the tag provides a detectable signal;
- after a sufficient time for said analyte and tagged analyte to bind to said cells, separating said cells from said liquid assay medium, substantially free of non-specifically bound tagged an analyte;
- and
- determining the amount of tagged analyte bound to said cells by means of said detectable signal, as compared to the amount of tagged analyte determined in an assay with a known amount of analyte.
1 more element
A method for determining an analyte capable of specifically binding to a cytosol receptor, said method comprising:
- combining in a buffered liquid nutrient assay medium supportive of cell viability a sample suspected of containing said analyte, cells having a specific intracellular receptor for said analyte and analyte tagged with a radioactive isotope;
- incubating said mixture at a temperature in the range of about 30.degree. to 45.degree. for a time sufficient for analyte and tagged analyte to bind to said cells;
- separating said cells substantially free of non-specifically bound tagged analyte from said liquid medium;
- disrupting said cells in liquid medium and separating the resulting cell debris from said liquid medium to provide a supernatant liquid;
- and
- determining the amount of radioisotope in the supernatant as compared to a supernatant obtained with a known amount of analyte.
3 more elements
Dependencies are inferred from the claim text.