Chromatin-nuclear envelope complex from rat liver: isolation and purification.
We describe a method for the isolation of a fraction of nuclear envelope (NE) from rat liver. The method includes mild treatment of pure nuclei with either endonuclease of DNase I under low ionic strength conditions in the presence of magnesium, which allows the nucleomeric organization of the chromatin (Ch) to be preserved. The NEs were purified by centrifugation in sucrose gradients followed by floatation in sucrose. No more than 3% of the Ch present in the purified Ch-NE complexes was due to the non specific adsorption of Ch to the NE. The main components of the complex (Ch and NE) retained their in situ ultrastructure. The complex consisted of 9--10% DNA, 3--4% RNA, about 63% protein and about 24% phospholipids.
D000818:Animals; D002458:Cell Fractionation / Q000379:methods; D002843:Chromatin / Q000302:isolation & purification* / Q000648:ultrastructure; D004247:DNA / Q000302:isolation & purification; D003851:Deoxyribonucleases / Q000378:metabolism; D005260:Female; D008099:Liver / Q000648:ultrastructure; D009685:Nuclear Envelope / Q000648:ultrastructure*; D009698:Nucleoproteins / Q000302:isolation & purification; D010743:Phospholipids / Q000302:isolation & purification; D012313:RNA / Q000302:isolation & purification; D051381:Rats
D002843:Chromatin; D009698:Nucleoproteins; D010743:Phospholipids; D012313:RNA; D004247:DNA; D003851:Deoxyribonucleases