Quantitative technique .
(Also referred to as electro-immunoassay or electro-immunodiffusion)
PRINCIPLE- Method of determining the concentration of antigen (Ag) in an unknown sample.
1. Make the agarose(purified agar) solution.
2. Add antiserum .
3. Pour the mixture into the glass plate .
4. Allow to solidify .
5. Place the glass plate in electrophoresis tank after puring the buffer.
6. Make some wells by puncturing with gel puncher without damaging the gel.
7. Load the antigens in the well.
8. Electrophorese the samples till the rockets are visible at 100 v.
9. Measure the height of the rocket and note the observation .
10. From the peak height concentration of antigen is determined.
(On the basis of their
electrophoretic mobility movement of antigens occur.)
The majority of the antibody-antigen precipitate is indeed at the head of the rocket.
Function- Used to differentiate between serum proteins (Albumin ,Fibrinogen, Globulin)
NOTE-Antigen concentration is directly proportinal to the height of the peak.