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Biokhimiia ; 59(12): 1852-9, 1994 Dec.
Article in Russian | MEDLINE | ID: mdl-7873684

ABSTRACT

DNA-dependent RNA polymerase IIB having a specific activity of 320 u./mg has been isolated from the term placenta homogenate using extraction performed at 4-6 degrees C in the presence of 75 mM ammonium sulfate and 1.5% nonidet P40, fractionation on DEAE-cellulose DE 23, desalting and heparin-agarose chromatography, resulting in 330-fold purification and a 18% yield. Technical details have been determined which are of crucial importance for reproducibility of affinity chromatography. The possibility of proteolysis of the IIc subunit during enzyme purification has been demonstrated.


Subject(s)
Placenta/enzymology , RNA Polymerase II/isolation & purification , Chromatography, Affinity , Chromatography, DEAE-Cellulose , Electrophoresis, Polyacrylamide Gel , Humans , Hydrolysis , RNA Polymerase II/metabolism , Reproducibility of Results
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