Extraction of DNA-binding proteins from cryogenically preserved cells.
We demonstrate that DNA-binding protein extracts can be effectively prepared directly from tissue culture cells preserved under liquid nitrogen without returning the cells to culture. We prepared DNA-binding protein extracts directly upon thawing of T47D, Jurkat and CAC-L153S cell lines after storage in liquid nitrogen for periods of up to one year. Our results show that DNA binding of a repressor of mouse mammary tumor virus transcription in these extracts is indistinguishable from binding activity in similar extracts prepared from cells maintained in culture.
Torrance
H
H
Department of Medicine, University of Ottawa Moses and Rose Loeb Institute for Medical Research, Ottawa Civic Hospital, Ontario, Canada.
MacLeod
H L
HL
Haché
R J
RJ
D001483:Base Sequence; D001665:Binding Sites; D002460:Cell Line; D015925:Cryopreservation; D004279:DNA, Viral / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000302:isolation & purification / Q000378:metabolism*; D008324:Mammary Tumor Virus, Mouse / Q000235:genetics*; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000302:isolation & purification / Q000378:metabolism; D012091:Repetitive Sequences, Nucleic Acid; D014158:Transcription, Genetic
D004279:DNA, Viral; D004268:DNA-Binding Proteins; D009687:Nuclear Proteins