Affinity chromatography of RNase inhibitor.
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.
Garcia
M A
MA
Department of Cellular and Structural Biology, University of Texas Health Science Center, San Antonio 78284, USA.
Klebe
R J
RJ
D000818:Animals; D002417:Cattle; D011232:Chemical Precipitation; D002846:Chromatography, Affinity / Q000379:methods*; D004791:Enzyme Inhibitors / Q000302:isolation & purification*; D011092:Polyethylene Glycols / Q000737:chemistry; D012260:Ribonucleases / Q000037:antagonists & inhibitors*
D004791:Enzyme Inhibitors; D011092:Polyethylene Glycols; D012260:Ribonucleases