A new high sensitive analytical micro-scale procedure for chromosomal proteins.
Utilizing male rat liver cells we describe a method for isolating and fractionating chromosomal proteins. About 99% of chromosomal proteins was dissociated using a three step dissociation procedure. DNA was removed by sedimentation and the histone fractions were separated from the non-histone chromosomal proteins by Bio Rex 70 chromatography. The non-histone chromosomal proteins were fractionated by micro-gradient electrophoresis on SDS-polyacrylamide gels, which proved to be superior to the electrophoretic procedures currently in use. The histones were further separated on polyacrylamide-SDS slab gels using a micro-two-dimensional electrophoretic system. The high resolution of these fractionation procedures greatly enhances the possibility of observing small changes in proteins which may play a role in gene regulation.
D000818:Animals; D002458:Cell Fractionation / Q000379:methods; D002467:Cell Nucleus / Q000032:analysis*; D002843:Chromatin / Q000032:analysis*; D002868:Chromosomal Proteins, Non-Histone / Q000302:isolation & purification*; D006657:Histones / Q000302:isolation & purification*; D008099:Liver / Q000648:ultrastructure; D008297:Male; D008832:Microchemistry / Q000379:methods; D051381:Rats
D002843:Chromatin; D002868:Chromosomal Proteins, Non-Histone; D006657:Histones