Chromatin prepared using micrococcal nuclease retains the enzyme activity which can be removed with cation-exchange resin AG 50 WX2.
Micrococcal nuclease used to digest nuclei remains bound to the chromatin in an inactive form. The enzyme activity can be restored in situ by addition of divalent cations such as Ca++ or Mg++ resulting in continued digestion of high molecular weight chromatin into smaller fragments as demonstrated by two dimensional gel electrophoresis of samples as chromatin and DNA in the first and second dimensions, respectively. The bound enzyme can be selectively removed from the chromatin by treatment with cation exchange resin AG 50 WX2 at low salt concentration without altering the electrophoretic mobility of the chromatin.
D000818:Animals; D002118:Calcium / Q000494:pharmacology; D002411:Cation Exchange Resins; D002467:Cell Nucleus / Q000201:enzymology; D002642:Chick Embryo; D002843:Chromatin / Q000201:enzymology*; D002852:Chromatography, Ion Exchange; D004247:DNA / Q000302:isolation & purification; D007700:Kinetics; D008099:Liver / Q000201:enzymology; D008274:Magnesium / Q000494:pharmacology; D008836:Micrococcal Nuclease / Q000302:isolation & purification / Q000378:metabolism*
D002411:Cation Exchange Resins; D002843:Chromatin; D004247:DNA; D008836:Micrococcal Nuclease; D008274:Magnesium; D002118:Calcium