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

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
(June 28, 1912 – September 30, 1978)

14. Examples Of Fluorescent Dyes
• Fluorescein Rhodamine
tetramethyl rhodamine is
othiocyanate (TRITC)
fluorescein isothiocyanate (FITC).

15. 1.Antibody tagged with flourescent dye
2.Antibody attached specifically to antigen
3.View specimen under exciting light
4.Flourscence microscope

16. 1. Direct immunofluorescence

17. 2. Indirect immunofluorescence:
•

19. 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

20. THANKYOU