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Assay of DNA-binding proteins with a dsDNA-coupled plate
- Source :
-
Clinical Biochemistry . Feb2006, Vol. 39 Issue 2, p167-175. 9p. - Publication Year :
- 2006
-
Abstract
- Abstract: Objectives: : This paper fabricated a cost-effective dsDNA-coupled plate (dcPlate) and applied it to measure the abundance and DNA-binding activity of a DNA-binding protein (DBP). Design and Methods: : The dcPlate was manufactured by covalently immobilizing an amino-modified oligonucleotide in wells of the plate coated with N-oxysuccinimide esters. The dcPlate was applied to measure the abundance of DNA-binding activity of a DBP in the same four steps, including protein incubation, primary antibody binding, enzyme-linked secondary antibody binding, and colorimetric development. Results: : The detections of three purified DBPs including NF-κB, AP1 and SP1, and HeLa cell nuclear extract and assays of DNA-binding activity of NF-κB p50 to five various DNA sequences demonstrated that dcPlate can be used to measure the abundance of DBPs quantitatively and assay DNA-binding activity of DBPs in high throughputs format. Conclusions: : The homemade cost-effective dcPlate provides a simple and versatile platform for studying DBPs. [Copyright &y& Elsevier]
- Subjects :
- *NUCLEIC acids
*CARRIER proteins
*ENZYME-linked immunosorbent assay
*SEX hormones
Subjects
Details
- Language :
- English
- ISSN :
- 00099120
- Volume :
- 39
- Issue :
- 2
- Database :
- Academic Search Index
- Journal :
- Clinical Biochemistry
- Publication Type :
- Academic Journal
- Accession number :
- 19771224
- Full Text :
- https://doi.org/10.1016/j.clinbiochem.2005.10.017