Immunoenzymatic

Frahm SO, Rudolph P, Dworeck C, et al. Immunoenzymatic detection of the new proliferation associated protein plOO by means of a cellular ELISA specific detection of cells in cell cycle phases S, G2 and M. J. Immunol. Methods 1999 223 147-153. 

Gillitzer, R., Berger, R., and Moll, H. (1990) A reliable method for simultaneous demonstration of two antigens using a novel combination of immunogold-silver staining and immunoenzymatic labeling. J. Histochem. Cytochem. 38, 307-313. 

Ishikawa, E., Imagawa, M., and Hashida, S. (1983b) Ultra sensitive enzyme immunoassay using fluoro-genic, luminogenic, radioactive and related substances and factors to limit the sensitivity. Proceedings of the 2nd International Symposium on Immunoenzymatic Technology. 

We have developed chemiluminescent immunoenzymatic assays for (3-ago-nist drugs in the 96-well-microtiter-plate format. Such competitive assays have been used for determination of clenbuterol and of the overall content of p-agonist drugs in the sample. They matched the standard requirements of precision and accuracy, and were more sensitive compared to the conventional colorimetric methods. Moreover, CL detection was very rapid, making these assays suitable for screening analysis. 

A collection of useful protocols for the simultaneous immunoenzyme staining of two or more tissue antigens is available on the websites http //www.protocol-online.org/prot/Immunology/ http //www.vectorlabs.com/Protocols/MLB.pdf and http //www.ihcworld.com/. The protocols given in this chapter are practiced in the authors laboratory. These methods fall into two main categories (a) simultaneous immunoenzymatic double staining, and (b) sequential immunoenzymatic double/ multiple staining. 

Basic protocol for simultaneous immunoenzymatic double staining using primary antibodies of two different species or two different IgG isotypes... 

Basic protocol for sequential immunoenzymatic double staining (Adapted from http //www.vectorlabs.com/ and http //www.ihcworld.com/ books/Dako Handbook.pdf)... 

Stripping Buffers for Sequential Immunoenzymatic Double Staining... 

Guesdon JL, Avrameas S. An immunoenzymatic method for measuring low concentrations of antigens by single radial diffusion. Immunochemistry 1974 11 595. 

Guesdon, J., Temynck, T., and Avrameas, S. (1979) The use of techniques avidin-biotin interaction in immunoenzymatic techniques. J. Histochem. Cytochem. 27, 1131-1139. 

Cordell, J. L., Falini, B., Erber, W. N., Ghosh, A. K., Abdulaziz, Z., MacDonald, S., Pulford, K. A. F., Stein, H., and Mason, D. Y. (1984) Immunoenzymatic labeling of monoclonal antibodies using immune complexes of alkaline phosphatase and monoclonal anti-alkaline-phosphatase (APAAP complexes). J. Histochem. Cytochem. 32, 219-229. 

Mason, D. Y. and Sammons, R. (1978) Alkaline phosphatase and peroxidase for double immunoenzymatic labeling of cellnlar constitnents. J. Clin. Pathol. 31, 454 60. 

Krenacs, T., Laszik, Z., and DobU, E. (1989) Application of immnnogold-silver staining and immunoenzymatic methods in mnltiple labeling of hnman pancreatic Langerhans islet cells. Acta Histochem. 85, 79-85. 

Martin, M.S., Hammann, A. Martin, F. (1986) Gut-associated lymphoid tissue and 1,2-dimethylhydrazine intestinal tumors in the rat a histological and immunoenzymatic study. Int. J. Cancer, 38, 75-80... 

Patulin (C6H704), is a compound, dangerous to health in humans which is found in the juice of damaged apples or grapefruits. The current method of measuring this compound resembles that of a immunoenzymatic ELISA test. A standard solution of pure patulin was prepared in water at a concentration of 1.54 g/1 just prior to use. 

Despite the fact that biological and predominantly immunoenzymatic methods have received increasing attention during the last decade for the determination of aflatoxins, chemical and immunochemical assays have to be preferred for their characteristics, including a lower limit of detection and high specificity. 

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