Abstract
Of the reagents used in reverse transcription-PCR (RT-PCR), RNase Inhibitor is the most costly on a per assay basis. A simple method for purification of RNase Inhibitor from bovine liver is described. Approximately 40 000 units of RNase Inhibitor can be purified to homogeneity from 400 g of bovine liver within two days using inexpensive reagents. The method described employs (a) facile procedures to rapidly obtain a clarified liver extract, (b) affinity chromatography of RNase Inhibitor on RNase-A-Sepharose, and (c) batchwise concentration of the purified protein with a molecular sieve resin.
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References
Blackburn P, Wilson G & Moore S (1977) J. Biol. Chem. 252:5904–5910
Burton L E & Fucci N P (1982) Int. J. Peptide Protein Res. 19:372–379
Holley R W, Apgar J, Doctor B P, Farrow J, Marini M A & Merrill S H (1961) J. Biol. Chem. 236:200-202
Klebe R J, Grant G M, Grant A M, Garcia MA, Giambernardi T A & Taylor G P (1996) BioTechniques (in press, November, 1996 issue)
Lee F S & Vallee B L (1993) Prog. Nucl. Acid Res. Mol. Biol. 44:1–30
March S C, Parikh I & Cuatrecasas P (1974) Anaylt. Biochem. 60:149–152
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Garcia, M.A., Klebe, R.J. Affinity chromatography of RNase Inhibitor. Mol Biol Rep 24, 231–233 (1997). https://doi.org/10.1023/A:1006818400674
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DOI: https://doi.org/10.1023/A:1006818400674