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These samples are now ready for SDS gel electrophoresis. The soybean trypsin inhibitor that is included acts as a carrier protein to facilitate quantitative precipitation. However, it also serves as an internal control for equivalent recovery and loading of each gradient fraction. Soybean trypsin inhibitor was chosen because its 21,000 molecular weight does not overlap with any of the translation initiation factor proteins (or their subunits) and thus will not interfere with evaluating whether a given protein (peptide) is present.

Drug Target 12, 125–134. , Carlson, C. , Beal, P. , and Pelletier, J. (2005). Inhibitory properties of nucleic acid-binding ligands on protein synthesis. FEBS Lett. 579, 79–89. Merrick, W. C. (1979a). Assays for eukaryotic protein synthesis. Methods Enzymol. 60, 108–123. Merrick, W. C. (1979b). Purification of protein synthesis initiation factors from rabbit reticulocytes. Methods Enzymol. 60, 101–108. Merrick, W. C. (1979c). Evidence that a single GTP is used in the formation of 80S initiation complexes.

Gao, H. , Merrick, W. , Hanamm, M. , and Pelletier, J. (2006b). Chlorissoclimides: New inhibitors of eukaryotic protein synthesis. RNA 12, 717–725. , and Schreier, M. H. (1979). Purification and characterization of seven initiation factors for mammalian protein synthesis. Methods Enzymol. 60, 136–165. , and Rhoads, R. E. (2001). Synthesis and properties of mRNAs containing the novel ‘‘anti-reverse’’ cap analogs 7-methyl(30 -O-methyl)GpppG and 7-methyl(30 deoxy)GpppG. RNA 7, 1486–1495. Svitkin, Y.

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