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.
Department of Medicine, University of Ottawa Moses and Rose Loeb Institute for Medical Research, Ottawa Civic Hospital, Ontario, Canada.
H
H
Torrance
H L
HL
MacLeod
R J
RJ
Haché
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