Test 2 extra credit 442-842.docx

In a radioreceptor essay we are determining the

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concentrations of different things in your sample. In a radioreceptor essay, we are determining the number of receptors present in the sample, which can then provide information on the ligand affinity. A radioimmunoassay is used to measure the amount of hormone in a given sample. This assay is based on the competition of labeled and unlabeled hormone for an antibody. For this test, an anti-leptin or anti- insulin antibody will be selected and added to separate test tubes in equal amounts. Then, labeled insulin or leptin will be added to each tube and allowed to incubate, so the two specifically bind to one another. After the incubation period, a serum sample (unlabeled hormone/antigen) is added. This causes competition for antibody binding between the hot and cold hormones in the test tube. As the concentration of cold antigen is increased, more hot-antigen is displaced, freeing the hot-antigen, and increasing the ratio of cold-antigen bound to the antibody, while reducing the ratio of labeled-antigen bound to the antibody to free labeled-antigen. The free, labeled-antigen can then be counted. This information is used to create a standard curve comparing the percent of bound hormone to the amount of hormone present. The standard curve can then be used to determine if a patients serum levels of hormone are low, adequate, or high, which may be reflected in their phenotype.
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