The Effect of Precipitation of the Complexes of Streptavidin with Biotinylated Proteins in Agar Gel

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Abstract

The effect of precipitation of complexes of streptavidin with biotinylated proteins under conditions of simple (according to Mancini) and double (according to Ouchterlony) radial diffusion in agar gel was studied. The position and form of precipitation lines depended primarily on the initial concentration of components and the degree of protein biotinylation. Free biotin, 1% SDS, and 6 M urea contained in the gel, as well as thermal denaturation of streptavidin inhibited the precipitate formation. Mannose, glucose, fucose, galactose, sucrose, and NaCl at high concentrations had no effect on biospecific precipitation. A model of interaction of streptavidin with biotinylated macromolecules is suggested, which accounts for the observed effect, and the prospects of practical application of the precipitation effect are discussed.

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