Sie sind auf Seite 1von 5

Ouchterlony double immunodiffusion

From Wikipedia, the free encyclopedia

Jump to: navigation, search This article does not cite any references or sources. Please help improve this article by adding citations to reliable sources. Unsourced material may be challenged and removed. (September 2011)

Picture of an Ouchterlony double immunodiffusion plate, after immunodiffusion has taken place. In this, titre value of an antigen is quantified. The central well has an antibody, and the surrounding wells have decreasing concentration of the corresponding antigen. Ouchterlony double immunodiffusion (also known as agar gel immunodiffusion or passive double immunodiffusion) is a simple, rather dated method which is still considered to be the gold standard for detection of extractable nuclear antigens. It is named after rjan Ouchterlony, the Swedish physician who invented the test.

[edit] Procedure
A gel plate is cut to form a series of holes ("wells") in the gel. A sample extract of interest (for example human cells harvested from tonsil tissue) is placed in one well, and sera or purified antibodies are placed in another well and the plate left for 48 hours to develop. During this time the antigens in the sample extract and the antibodies each diffuse out of their respective wells. Where the two diffusion fronts meet, if any of the antibodies recognize any of the antigens, they will bind to the antigens and form what is known as an immune complex. This immune complex precipitates in the gel to give a thin white line, which is a visual signature of antigen recognition. The method can be conducted in parallel with multiple wells filled with different antigen mixtures and multiple wells with different antibodies or mixtures of antibodies, and antigenantibody reactivity can be seen by observing between which wells the precipitate is observed. When more than one well is used there are many possible outcomes based on the reactivity of the antigen and antibody selected. The zone of equivalence lines may give a full identity (i.e. a continuous line), partial identity (i.e. a continuous line with a spur at one end), or a non-identity (i.e. the two lines cross completely).

[edit] Theory

Precipitation occurs with most antigens because the antigen is multivalent (i.e. has several antigenic determinants per molecule to which antibodies can bind). Antibodies have at least two antigen binding sites (and in the case of Immunoglobulin M there is a multimeric complex with up to 10 antigen binding sites), thus large aggregates or gel-like lattices of antigen and antibody are formed. Experimentally, an increasing amount of antigen is added to a constant amount of antibody in solution. Initially at low antigen concentration, all of the antigen is contained in the precipitate. This is called the antibody-excess zone (i.e. prozone phenomenon). As more antigen is added, the amount of protein precipitated increases until the antigen/antibody molecules are at an optimal ratio. This is known as the zone of equivalence or equivalence point. When the amount of antigen in solution exceeds the amount of antibody, the amount of precipitation will decrease. This is known as the antigen excess zone.

[edit] External links

Simulator at University of California, Irvine Overview at University of Maryland [hide]

v t e

Medical test: Immunologic techniques and tests (CPT 8600086849)

Chromatin immunoprecipitation Immunodiffusion (Ouchterlony double Immunoprecipitation immunodiffusion, Radial immunodiffusion, Immunoelectrophoresis, Counterimmunoelectrophoresis) ELISA ELISPOT Enzyme Multiplied Immunoassay Technique RAST test Immunoassay Radioimmunoassay Radiobinding assay Immunofluorescence Hemagglutination/Hemagglutinin Agglutination (Coombs test) Latex fixation test Nephelometry Complement fixation test Immunocytochemistry Other Immunohistochemistry (Direct fluorescent antibody) Epitope mapping Skin allergy test Patch test C-reactive protein Procalcitonin Inflammation Total complement activity MELISA

Immunologic techniques and tests serology/ diagnostic immunology


CBC (lymphocyte count) cell/phys/auag/auab/comp, imdf/ipig/hyps/tumr igrc

proc, drug(L3/4)

Retrieved from " 5699" View page ratings Rate this page What's this? Trustworthy Objective Complete Well-written I am highly knowledgeable about this topic (optional) I have a relevant college/university degree It is part of my profession It is a deep personal passion The source of my knowledge is not listed here I would like to help improve Wikipedia, send me an e-mail (optional) We will send you a confirmation e-mail. We will not share your e-mail address with outside parties as per our feedback privacy statement. Submit ratings Saved successfully Your ratings have not been submitted yet Your ratings have expired Please reevaluate this page and submit new ratings. An error has occurred. Please try again later. Thanks! Your ratings have been saved. Please take a moment to complete a short survey. Start surveyMaybe later Thanks! Your ratings have been saved. Do you want to create an account? An account will help you track your edits, get involved in discussions, and be a part of the community. Create an accountorLog inMaybe later Thanks! Your ratings have been saved. Did you know that you can edit this page? Edit this pageMaybe later


Biological techniques and tools Immunologic tests

Hidden categories:

Articles lacking sources from September 2011 All articles lacking sources

Personal tools

Create account Log in


Article Talk

Variants Views Actions Search


Read Edit View history


Main page Contents Featured content Current events Random article Donate to Wikipedia


Help About Wikipedia


Community portal Recent changes Contact Wikipedia

What links here Related changes Upload file Special pages Permanent link Cite this page Rate this page


Create a book Download as PDF Printable version


Deutsch Espaol Romn This page was last modified on 19 November 2011 at 09:27. Text is available under the Creative Commons Attribution-ShareAlike License; additional terms may apply. See Terms of use for details. Wikipedia is a registered trademark of the Wikimedia Foundation, Inc., a non-profit organization. Contact us Privacy policy About Wikipedia Disclaimers Mobile view