Efficacy of RNase inhibitors during brain polysome isolation.
We have investigated the efficiency of heparin, polyvinyl sulfate and yeast RNA (as competitive RNase inhibitors), liver extract (as crude preparation of liver RNase inhibitors) and DEPC (as irreversible non-competitive inhibitor) for the preparation of rat brain polysomes. Sucrose gradient sedimentation profiles, obtained from PMS, were used to determine the optimal concentration of each inhibitor. Diethylpyrocarbonate, whatever the composition of isolation buffer, was found detrimental for brain polysomes. Most of the other inhibitors where found useless or even harmful. A slight positive effect was observed with heparin 0.75 mg/mL both for total yield and sedimentation pattern. It is concluded that the utilisation of most of the widely used RNase inhibitors is of questionnable effectiveness for brain polysome preparation.
D000818:Animals; D001921:Brain / Q000648:ultrastructure*; D002458:Cell Fractionation / Q000379:methods*; D004047:Diethyl Pyrocarbonate / Q000494:pharmacology; D006493:Heparin / Q000494:pharmacology; D008110:Liver Extracts / Q000494:pharmacology; D011132:Polyribosomes / Q000187:drug effects*; D011145:Polyvinyls / Q000494:pharmacology; D012313:RNA / Q000302:isolation & purification / Q000494:pharmacology; D051381:Rats; D012260:Ribonucleases / Q000037:antagonists & inhibitors*; D012441:Saccharomyces cerevisiae / Q000032:analysis
D008110:Liver Extracts; D011145:Polyvinyls; C019050:polyvinyl sulfate; D012313:RNA; D006493:Heparin; D012260:Ribonucleases; D004047:Diethyl Pyrocarbonate