Antibody method

In the double antibody method of separation, a second antibody, produced by injecting the first antibody into another animal, is utilized. This antibody is used to combine with and form an insoluble complex with the first antibody. After... 

Nakane, P.K. (1975) Recent progress in the peroxidase-labeled antibody method. Ann. N.Y. Acad. Sci. 254, 203. 

Albert H. Coons (Fig. 1.2) was the first who attached a fluorescent dye (fluorescein isocyanate) to an antibody and used this antibody to localize its respective antigen in a tissue section. The concept of putting a visible label on an antibody molecule appeared both bold and original. His initial results were described in two brief papers in the early 1940s (Coons et al. 1941,1942), but the research was halted while he joined the army and spent the next 4 years in the South Pacific. His later studies (Coons and Kaplan 1950) contributed immensely to the use of the fluorescent antibody method in a wide variety of experimental settings. In our time, the use of antibodies to detect and localize individual or multiple antigens in situ has developed into a powerful research tool in almost every field of biomedical research (http //books.nap.edu/html/biomems/acoons.pdf). 

Figure 26 Production of abzymes using the anti-idiotypic antibodies method.

Hsu, S. M., Cossman, J. C., and Jaffe, E. S. (1983) A comparison of ABC, unlabeled antibody and conjugated antibody methods with monoclonal and polyclonal antibodies—an examination of germinal center of tonsils. Am. J. Clin. Pathol. 80, 429 35. 

Nakane, P. K. (1968) Simultaneous localization of multiple tissue antigens using the peroxidase-labeled antibody method a study on pituitary glands of the rat. J. Histochem. Cytochem. 16, 557-560. 

Vroman has shown by antibody methods that plasma interactions with solid surfaces result in a hierarchial adsorption process 98). The high concentration proteins dominate the surface at short times due to the higher collision rates. As time passes... 

Because of the superior morphology provided by formalin-fixed paraffin-embedded tissues, this has become the medium of choice for most clinical and research studies. The peroxidase-labeled antibody method, introduced in 1968, was the first practical application of antibodies to paraffin-embedded tissues and overcame some of the limitations of earlier fluorescence antibody methods (1). These pioneering studies using enzyme labels instead of fluorescent dyes opened the door to the development of modern methods of immuohistochemistry. 

Krensky, A. M. and Nelson, P. J. (1997) Expression of the chemokine RANTES and production of monoclonal antibodies. Methods Enzymol. 287,162-174. 

Hwang WYK, Foote J. Immunogenicity of engineered antibodies. Methods 2005 36 3-10. 

There are two methods the preformed radiometal-chelate method and the indirect chelator—antibody method. Various antibodies are labelled by the latter, where the bifunctional chelating agent is initially conjugated to a macromolecule, which is then allowed to react with a metal ion, to form a metal-chelate-macromolecule complex. Due to the presence of the chelating agent, the biological properties of the labeled protein may be altered and must be assessed before clinical use. 

Dunbar, B.S., Schwoebel, E.D. (1990) Preparation of Polyclonal Antibodies, Methods Enzymol. 182, 663-670. 

Polyclonal antibody production is the most primitive method and has several significant limitations (6). This method inherently yields a complex mixture of antibody molecules from the immune response after hapten hypeiimmunization with no attempt to purify specific IgG molecules. Efficiency and cost-effectiveness are hallmarks of polyclonal antibody production however, accurate characterization of this mixture is difficult. Furthermore, X-ray crystallography to examine structure-function relationships and affiiuty maturation to optimize the antibody is impossible with the polyclonal antibody method. 

Shokat KM, Schultz PG. Catalytic antibodies. Methods Enzymol. 1991 203 327-351. 

Use of the Double-Antibody Method to Separate Antibody Bound from Free Ligand in Radioimmunoassay... 

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