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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
Foi desenvolvido um ensaio quantitativo de ligação de ADN à base de ELISA para medir as interacções com o factor de transcrição do ADN. Alta especificidade para a proteína RUNX2 foi conseguida com um oligonucleótido de ADN de reconhecimento de consenso e de anticorpo monoclonal específico. Detecção Colorimétrico com uma reacção do substrato, o anticorpo acoplado a enzima foi monitorizada em tempo real.
