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A Free-Labeled Method for DNA-Binding Protein Detection Using a Double-Stranded DNA Microarray

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Abstract:

We have developed a new type of double-stranded DNA microarray to perform detection of sequence-specific DNA-binding proteins. The DNA-binding site of a DNA-binding protein is divided into two fragments. One fragment was immobilized on an aldehyde-coated glass microscope slide surface via chemical bonds. The other fragment was labeled with a fluorescent molecule. When using this kind of double-stranded DNA microarray, the labeled DNA fragment was pre-incubated with detection sample for 5 to 10 minutes and then hybridized with the microarray. In our experiment, six different concentrations of Nuclear Factor -B P50 homodimer in detection samples were tested. The microarray fluorescence intensity was obtained and the relationship between the intensity and the protein concentration was calculated. The detection results suggested that this free-labeled detection system could have the ability to be used in research and medical diagnosis and for high-throughput screening of drugs targeted to DNA-binding proteins.

Keywords: DNA-BINDING PROTEIN; DOUBLE-STRANDED DNA MICROARRAY; DSDNA; FLUORESCENCE; FREE-LABELED DETECTION

Document Type: Research Article

DOI: http://dx.doi.org/10.1166/jnn.2005.206

Publication date: August 1, 2005

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  • Journal for Nanoscience and Nanotechnology (JNN) is an international and multidisciplinary peer-reviewed journal with a wide-ranging coverage, consolidating research activities in all areas of nanoscience and nanotechnology into a single and unique reference source. JNN is the first cross-disciplinary journal to publish original full research articles, rapid communications of important new scientific and technological findings, timely state-of-the-art reviews with author's photo and short biography, and current research news encompassing the fundamental and applied research in all disciplines of science, engineering and medicine.
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