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RIA, IMMUNOFLURESCENCE ASSAY
Radioimmunoassay (RIA) is an in vitro assay technique introduced in 1960 that uses antibodies to very sensitively measure antigen concentrations. It involves the separation of a protein from a mixture using the specificity of antibody-antigen binding, followed by quantitation using radioactivity. RIA utilizes competitive binding reactions and the measurement of radio emissions. Immunofluorescence also uses fluorescent-labeled antibodies but to detect specific target antigens. It emits greenish fluorescence under UV light that can be viewed under a fluorescence microscope. Both techniques have various medical applications like cancer detection, drug screening, and research.
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RIA, IMMUNOFLURESCENCE ASSAY
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- 2. Radioimmunoassay (RIA) • Radioimmunoassay(RIA) is a very sensitive in vitro assay technique used to measure concentrations of antigens by use of antibodies. HISTORY • The technique was introduced in 1960 by Berson and Yalow as an assay for the concentration of insulin in plasma.
- 3. Dr. Rosalyn YalowDr. Solomon Berson
- 4. “The world cannotafford the loss of the talents of half its people if we are to solve the many problems which beset us.”
- 5. Labels In Immunoassays •Immunoassays require the use of labeled materials in order to measure the amount of antigen or antibody present. • A label is a molecule that will react as part of the assay, and in doing so produce a signal that can be measured in the solution.
- 6. PRINCIPLE • Radioimmunoassay (RIA)involves the separation of a protein (from a mixture) using the specificity of antibody - antigen binding and quantitation using radioactivity.
- 7. THE TECHNIQUE • ThePrinciple of Radioimmunoassay (RIA) • An immune reaction i.e. antigen, antibody binding. • A competitive binding or competitive displacement reaction. (It gives specificity) • Measurement of radio emission. (It gives sensitivity) • A mixture is prepared of radioactive antigen antibodies against that antigen. • Known amounts of unlabeled ("cold") antigen .
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- 11. Uses for Radioimmunoassay •Narcotics (drug) detection • Blood bank screening for the hepatitis a highly contagious condition virus • Early cancer detection • Measurement of growth hormone levels • Tracking of the Leukemia virus • Research with brain chemicals called neurotransmitters.
- 12. IMMUNOFLUORESCENCE • Immunofluorescence :Immunofluorescence is a powerful technique that utilizes fluorescent- labeled antibodies to detect specific target antigens • Fluorescein is a dye which emits greenish fluorescence under UV light. It can be tagged to immunoglobulin molecules.
- 13. Albert Hewett Coons (June28, 1912 – September 30, 1978)
- 14. Examples Of FluorescentDyes • Fluorescein Rhodamine tetramethyl rhodamine is othiocyanate (TRITC) fluorescein isothiocyanate (FITC).
- 15. 1.Antibody tagged withflourescent dye 2.Antibody attached specifically to antigen 3.View specimen under exciting light 4.Flourscence microscope
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- 19. Applications • Detect thepresence and amount of antinuclear antibody (ANA) • Detect particular proteins • Studying DNA sequences on chromosomes • The deeper layers of a cell or tissue
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