Purification of DNA-binding proteins using tandem DNA- affinity column
نویسندگان
چکیده
منابع مشابه
Purification of sequence-specific DNA-binding proteins by affinity chromatography.
The affinity chromatography procedure described in this unit uses DNA containing specific recognition sites for the desired protein that has been covalently linked to a solid support. Preparation of a DNA affinity resin, including cyanogen bromide (CNBr) activation of the agarose support, is described, and an alternate protocol provides a method to couple DNA to commercially available CNBr-acti...
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Characterization of proteins binding to the promoters of eukaryotic genes has proved essential for understanding the transcriptional regulation of viral and cellular genes. Because of the low abundance of these proteins in the cell, conventional purification of these molecules has been laborious. In contrast, sequencespecific DNA affinity chromatography has greatly faciliated rapid isolation an...
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A nucleic acid affinity matrix containing a short oligodeoxynucleotide ligand has been prepared as an example of a material which can be used for the rapid and effective isolation of sequence specific DNA binding proteins. Two complementary oligodeoxynucleotides have been employed, one of which contains a small 5'-spacer arm with a terminal thiol group. Using this terminal thiol group, the liga...
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A 66-kD Alu-DNA-repeat binding protein was identified in human somatic cell nucleoplasm. Gel shift assay, southwestern blotting, and affinity purification on DNA attached to a carrier were used. A 60-kD protein copurified with the 66-kD protein during affinity purification, probably due to protein--protein interactions. The gel shift assay reveals multiple complexes with exponential dependence ...
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ژورنال
عنوان ژورنال: Nucleic Acids Research
سال: 1993
ISSN: 0305-1048,1362-4962
DOI: 10.1093/nar/21.4.1045