Ouchterlony method

In the Ouchterlony method, the double diffusion experiment is carried out with a central well containing the antibody or antibody mixture, surrounded by a circular... 

In 3 cases of Barta and Tichy (B3) with protein leakage into the gastric juice, serum proteins were identified by the Ouchterlony method. Hollander and Horowitz (H13), using this diffusion technique in canine gastric juice, demonstrated presence of serum albumin in canine gastric juice. 

Double immunodiffusion in two dimensions is a widely used immunotechnique and is known as the Ouchterlony method. It allows direct comparison of two or more test materials and provides a simple and direct method for determining whether the antigens in the test specimens are identical, cross reactive, or nonidentical. 

As stated in the introduction, immunoelectrophoresis (abbreviated ImEl) is a two-stage method. The electrophoretic run is comparatively simple and will be dealt with in a following chapter. The immunochemical reaction in agar gel is more complex, and this makes it necessary to examine the advantages of gel difEusion methods as well as their limitations. Many investigators use the Ouchterlony method, preceding or parallel to ImEl. This method will become still more important if present attempts at quantitation gain definite shape. 

Paper/starch first analysis of nucleotides and chemical synthesis products inherited changes in proteins + antisera- antigen recognition (Ouchterlony method)... 

Since the end of 1950s, the Ouchterlony method (Ouchterlony 1953) has been widely used with a large variety of antigen-antibody systems bacteria, moulds, yeasts and other organic material with antigenic properties. The fact that the Ouchterlony method is of low cost and is easy to use and to reproduce explains the long-term use of this technique. 

In order to investigate the active sites of these proteins, laccases I and III were subjected to ESR (electron spin resonance) spectroscopic analysis. The ESR spectra shown in Figure 5 indicate clear differences in peaks 2 and 6 which support the concept that the copper atoms in laccases I and III have different conformations in each molecule. Furthermore, immunological similarity between laccases I and III was also investigated. Antibody specific for laccase III was prepared from rabbit serum by conventional methods. When applied to Ouchterlony diffusion plates containing laccase I, no precipitation lines developed (Figure 6). This result showed that there were no conserved epitopes on the surfaces laccases I and III. 

Tanahashi et al. (1968) compared the immunological properties of bovine, water buffalo, ovine, caprine, porcine, guinea pig, and human a-lactalbumins by the method of Oudin. They found that the nonruminant a-lactalbumins do not react with antisera to the bovine protein. This is in accordance with our experience (K. Bell and H. A. McKenzie) using the Ouchterlony and immunoelectrophoretic methods. However, Sakar et al. (1971) found that while bovine, water buffalo, and caprine a-lactalbumins exhibit extensive cross-reaction in the Ouchterlony test, these ruminant a-lactalbumins may be differentiated quanti-... 

The most often used procedures for evaluating the specificity of antibody preparations are double diffusion methods developed by Ouchterlony and immunoelectrophoretic methods developed by Grabor and Williams. For a discussion of variations of these techniques as well as other useful procedures the reader is referred elsewhere (7-9). 

Ouchterlony technique. Double-radial immunodiffusion for the detection of precipitating autoantibodies against extractable nuclear antigens . Method of high diagnostic specificity but low sensitivity for diagnosis of autoimmune rheumatic diseases. 

Czerkinsky, C.C., Nilsson,L.A., Nygren,H., Ouchterlony, O., and Tarkowski, A. (1983) A solid phase enzyme linked immunospot (ELISPOT) assay for enumeration of specific antibody secreting cells. Journal of Immunological Methods, 65, 109 121. 

Czerkinsky, C.C., Andersson, G., Ekre, H.P., Nilsson, L.A., Klareskog, L., and Ouchterlony, O. (1988) Reverse ELISPOT assay for clonal analysis of cytokine production. I. Enumeration of gamma interferon secreting cells. Journal of Immunolog ical Methods, 110, 29 36. 

Ouchterlony, O., In vitro method for testing the toxin-producing capacity of diphtheria bacteria. Acta Pathol. Microbiol. Scand. 25, 186 (1948). 

Egg-white contains several distinct antigens such as ovalbumin, ovoglobulin, ovomucoid, ovomucin, conalbumin and lysozyme [283, 663] and immunochemical techniques have been used to characterize them [131, 338, 674]. Kaminski and Ouchterlony [341] used a combination of precipitation methods and electrophoresis to show 10 antigenically distinct fractions in egg-white. Tiselius and Eriksson-Quensel [621] had earlier indicated that ovalbumin is not electrophoretically homogeneous and subsequent electrophoretic studies showed that ovalbumin and conalbumin each separated into 2 fractions and ovoglobulin separated into 3 [118, 179, 180, 419, 424]. Other studies show that there may even be more protein sub-units in egg-white [48, 129, 144,184, 358, 359, 516]. 

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