Objective: To learn the technique of Ouchterlony Double Diffusion (ODD).
Principle: Interaction between antigen (Ag) and antibody (Ab) at the molecular level forms the basis for several techniques that are useful in modern day scientific studies and in routine clinical diagnosis. These techniques are either based on the use of labeled reagents, a tracer or immunoprecipitation. Ouchterlony double diffusion (ODD) or double immunodiffusion technique is one of the simplest techniques extensively used to check antisera for the presence of antibodies for a particular Ag and to determine its titre.
In ODD assays, solutions of Ag and Ab are placed in adjacent wells cut in agarose gel and are allowed to diffuse radially. The Ag and Ab concentrations are relatively higher near their respective wells. As they diffuse farther from the wells, their concentration decreases. An antigen will react with its specific antibody to form an Ag-Ab complex. At one point their concentrations become equivalent and the Ag-Ab complex precipitates to form a precipitin line.
Components of the kit 10 reactions:
Materials Quantity Store
Agarose 1gm 4°C
10 X Assay Buffer 10ml 4°C
Antigen 0.2ml -20°C
Antiserum 0.4ml -20°C
Glass plates 4 Nos. RT
Gel puncher 1 No RT
Template 2 Nos. RT