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Article: Real-time PCR Assay for Ultrasensitive Quantification of DNA-Binding Proteins
- Article from:
- Clinical Chemistry
- Article date:
- April 1, 2007
- Author:
CopyrightCopyright American Association for Clinical Chemistry Apr 2007. Provided by ProQuest LLC. (Hide copyright information)
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Background: The specific binding of proteins to DNA is a key step for many cellular activities, such as transcription regulation, DNA replication, recombination, repair, and restriction. The detection of DNA-binding proteins, as well as the identification of specific binding sites, is therefore important to understand gene expression mechanisms and cellular function. We describe an ultrasensitive method for quantification of DNA-binding proteins.
Methods: We combined the common exonuclease III (ExoIII) footprinting assay and real-time PCR for quantification of DNA-binding proteins, for an assay that does not require antibodies against the target proteins. Double-strand DNA probes were ...