Abstract

Antibodies can be used as highly specific reagents to detect, identify, or purify proteins. Polyvalent antisera directed against transport proteins have been used in a limited number of circumstances to investigate epithelial function. They have a disadvantage in that they are a mixed reagent and require a highly purified antigen in order to be specific. Monoclonal antibodies can be generated with the hybridoma technique and offer the advantage of being a pure antibody derived from a single antibody-producing cell directed at a single antigenic determinant. The methods of generation and screening are described. Monoclonal antibodies have been most frequently used to characterize membrane transport proteins such as Na-K-ATPase and H-K-ATPase. They provide a useful tool for microscopic localization and for probing synthesis, assembly, and insertion of specific proteins. The antigenic composition of nephron segments and polarization of epithelial cells have been studied with these methods. They have a great potential use in studies employing membrane vesicles. Monoclonal antibody technology offers the opportunity to correlate the biochemical, functional, and morphological characteristics of renal tubular epithelial transport.

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