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Nucleic Acids Research, 1979, Vol. 6, No. 6 2179-2191
© 1979

Articles

Isolation and characterization of rabbit anti-m7G-5'-P antibodies of high apparent affinity

R. Dickinson Meredith and Bernard F. Erlanger

Department of Microbiology, Cancer Center/Institute of Cancer Research, Columbia University, College of Physicians and Surgeons New York, NY 10032, USA

Received March 27, 1979. Antibodies highly specific for intact pm7G (7-methylguanosine-5'-mono-phosphate) were induced by immunization of rabbits with a pm7G-BSA conjugate. Since the nucleotide is six-fold more stable than m7G (7-methylguanosine) to alkali-catalyzed fission of the imidazole ring, it is a more desirable antigen for obtaining antibodies capable of binding caps on eukaryotic mRNA. UV spectra demonstrated that the nucleotide in the conjugate was predominantly the intact form. Competition radioimmunoassay showed 1) high apparent affinities for pm7G, on the order of 10–8 M, 2) low competition by the ringopened form of the homologous hapten (*pm7G), and by m7G, 3) little or no competition by AMP, GMP, CMP, UMP or m6A, and 4) high apparent affinities for m7GpppAm, m7GpppN6mAm, m7GpppGm, m7GpppA.


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