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Nucleic Acids Research, 1977, Vol. 4, No. 1 71-84
© 1977

Articles

Isolation of specific messenger RNA by adsorption of polysomes to matrix-bound antibody

Gunther Schutz, Shalom Kieval, Bernd Groner, Albrecht E. Sippel, David T. Kurtz and Philip Feigelson

Institute of Cancer Research and Department of Biochemistry, Columbia University New York, NY 10025, USAMax Planck Institut fur Molekulare Genetik Berlin, GFR

Received September 13, 1976. A procedure is presented for the purification of specific mRNAs, which exploits the ability of antibodies prepared against a native protein to bind to the nascent polypeptide on the polysome. Rather than precipitating these soluble antibody-polysome complexes with anti-antibody, which canlead to nonspecific trapping of polysomes, we have linked the anti-antibody to an insoluble matrix. Thus, the antibody-polysome complex binds to the anti-antibody support and nonspecific polysomes can easily be removed by several washes. We have found para -aminobenzyl cellulose (PAB cellulose), to be a suitable matrix for this purpose. This support can bind large quantities of anti-antibody and it displayed no detectable nonspecific affinity for polysomes or RNA. Using this procedure, we have obtained an apparently homogeneous preparation of ovalbumin mRNA.


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