Techniques of Precipitation

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Introduction:

Precipitation is a process in which soluble antigen binds to corresponding antibody to form a soluble precipitate (in contrast to agglutination where particulate antigen binds to corresponding antibody to form visible clumps).

Techniques of Precipitation

  1. Single diffusion in one dimension:
    • Ab is incorporated in agar gel in a test tube & Ag solution is layered over it
    • Ag diffuses downward through the agar gel forming a line of precipitation
    • Ex.: Oudin procedure.
  2. Double diffusion in one dimension:
    • Ab is incorporated in agar gel above which a column of plain agar is placed
    • The Ag is layered over it
    • The Ag & Ab move towards each other through the intervening column of plain agar & form the precipitate
    • Ex.: Oakley-Fulthorpe procedure.
  3. Single diffusion in double dimension:
    • Here the antisera is incorporated in a gel & poured on a flat surface
    • Wells are cut on the surface to which Ag is added
    • It diffuses radially from the well & forms ring shaped bands of precipitation concentrically around the well
    • Ex.: Radial immunodiffusion.
  4. Double diffusion in 2 dimensions:
    • Agar gel is poured on a slide & wells are cut
    • Antiserum is put into the central well
    • Different Ags are put in the surrounding wells
    • Ex.: Ouchterlony procedure & Elek’s gel precipitation.

Summary:

Diffusion

Dimension

Example

1

1

Oudin

2

1

Oakley-Fulthorpe

1

2

Radial immunodiffusion

2

2

Ouchterlony procedure

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