Nonradioactive assay for sequence-specific DNA binding proteins.
A nonradioactive functional assay was developed to quantitate DNA binding proteins. The assay was designed to allow the use of 96-well microplates for high sample throughput. We show that the assay can measure sequence-specific DNA binding of purified proteins as well as DNA binding activity present in whole cell extracts. By using a temperature-sensitive DNA binding protein, we demonstrate that DNA binding as measured by this novel assay correlates with the biological function of the protein.
Gubler
M L
ML
Department of Oncology, Roche Research Center, Hoffmann-LaRoche Inc., Nutley, NJ 07110, USA.
Abarzúa
P
P
D000818:Animals; D000888:Antibodies, Anti-Idiotypic; D001483:Base Sequence; D001710:Biotin; D002457:Cell Extracts / Q000235:genetics; D002460:Cell Line; D004268:DNA-Binding Proteins / Q000032:analysis*; D004797:Enzyme-Linked Immunosorbent Assay; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D012680:Sensitivity and Specificity; D016159:Tumor Suppressor Protein p53 / Q000032:analysis
D000888:Antibodies, Anti-Idiotypic; D002457:Cell Extracts; D004268:DNA-Binding Proteins; D015345:Oligonucleotide Probes; D016159:Tumor Suppressor Protein p53; D001710:Biotin