Efficacy of RNase inhibitors during brain polysome isolation |
| |
Authors: | Didier Gauthier M R Ven Murthy |
| |
Institution: | (1) Département de biochimie, Faculté de médecine, Université Laval, G1K 7P4 Québec, QC, Canada;(2) Present address: Département de chimie et biochimie, Université de Moncton, E1A 3E9 Moncton, N.-B., Canada |
| |
Abstract: | 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.Abbreviations DEPC
diethylpyrocarbonate
- PVS
polyvinyl sulfate
- PMS
post-mitochondrial supernatant
- HSS
high-speed supernatant |
| |
Keywords: | Polysomes RNase inhibitors |
本文献已被 SpringerLink 等数据库收录! |
|