RIA, IMMUNOFLURESCENCE ASSAY

RIA, IMMUNOFLURESCENCE

1. RADIOIMMUNO ASSAY IMMUNOFLUORESCENCE AMAL GEORGE SBS MGU

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 Yalow Dr. Solomon Berson

4. “The world cannot afford 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 The Principle 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 .

8. Procedure

9. 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 L eukemia virus Research with brain chemicals called neurotransmitters.

10. IMMUNOFLUORESCENCE I mmunofluorescence : 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.

11. Albert Hewett Coons (June 28, 1912 – September 30, 1978)

12. Examples Of Fluorescent Dyes Fluorescein Rhodamine tetramethyl   rhodamine   isothiocyanate  (TRITC) fluorescein  isothiocyanate  (FITC).

13. Antibody tagged with flourescent dye Antibody attached specifically to antigen View specimen under exciting light Flourscence microscope

14. Direct immunofluorescence

15. Indirect immunofluorescence:

16. Applications Detect the presence 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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