title: Preparation of ribonucleoprotein-enriched nucleoplasmic extracts from Drosophila cells.
abstract: Here we describe how to prepare nucleoplasmic extracts from suspension cultures of Drosophila S2 (Schneider Line 2) cells. Harvested cells are washed in phosphate-buffered saline supplemented with MgCl2, resuspended in hypotonic buffer, and homogenized. Nuclei are isolated by centrifugation and then sonicated. The nuclear sonicate is placed on a 30% sucrose cushion and sedimented. The soluble nuclear ribonucleoprotein (RNP) complexes remain in the supernatant and the nuclear membrane fragments and chromatin pellet through the sucrose cushion.
authors.forename: Donald C
authors.initials: DC
authors.lastname: Rio
mesh_terms: D000818:Animals; D002467:Cell Nucleus / Q000378:metabolism*; D003593:Cytoplasm / Q000378:metabolism*; D004330:Drosophila / Q000166:cytology*; D012261:Ribonucleoproteins / Q000378:metabolism*
chemical_list: D012261:Ribonucleoproteins