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1Greenebaum Cancer Center,University of Maryland School of Medicine, 2Program in Molecular Medicine,University of Maryland School of Medicine, 3Department of Biochemistry & Molecular Biology,University of Maryland School of Medicine, 4Department of Pharmacology & Experimental Therapeutics,University of Maryland School of Medicine, 5Department of Pathology and Biochemistry & Molecular Biology,University of Maryland School of Medicine
Chapters
- 00:05Title
- 01:49Cell Culture and Nuclear Protein Isolation
- 02:46Preparation of 96-well Plates by Incubation with Nuclear Extracts
- 03:58Measurement of RUNX2 DNA Binding
- 05:36Results: Inhibition of RUNX2 DNA Binding
- 06:22Conclusion
Vi utvecklade en kvantitativ DNA-bindning ELISA-baserad analys för att mäta transkriptionsfaktor interaktion med DNA. Hög specificitet för Runx2 protein uppnåddes med en konsensus-DNA-erkännande oligonukleotiden och specifik monoklonal antikropp. Kolorimetrisk detektion med en enzym-kopplad antikropp substratreaktionen övervakades i realtid.
