A simplified method for the preparation of transcriptionally active liver nuclear extracts.
We have developed a simplified method for the preparation of liver nuclear extracts to study gene regulation and protein-DNA interactions. This protocol uses conventional laboratory equipment and standard reagents. The liver tissue is homogenized in a low-salt solution at physiological molarity with subsequent adjustment of the molarity and purification of nuclei by density sedimentation. The nuclear extracts are transcriptionally active in a validated cell-free transcription assay and contain functional DNA-binding proteins. This protocol results in the rapid preparation of highly reproducible and active liver nuclear extracts.
Hattori
M
M
Department of Medicine, University of California, San Diego, La Jolla 92093.
Tugores
A
A
Veloz
L
L
Karin
M
M
Brenner
D A
DA
D000818:Animals; D001483:Base Sequence; D002467:Cell Nucleus / Q000378:metabolism; D002474:Cell-Free System; D004247:DNA / Q000235:genetics; D004268:DNA-Binding Proteins / Q000235:genetics; D005786:Gene Expression Regulation; D008099:Liver / Q000166:cytology / Q000378:metabolism*; D008969:Molecular Sequence Data; D010957:Plasmids; D051381:Rats; D013698:Templates, Genetic; D014158:Transcription, Genetic
D004268:DNA-Binding Proteins; D004247:DNA