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En kvantitativ analyse for å studere Protein: DNA interaksjoner, Discover Transcriptional regulatorer Gene Expression, og identifisere nye anti-tumor Agenter
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JoVE Journal Biology
A Quantitative Assay to Study Protein:DNA Interactions, Discover Transcriptional Regulators of Gene Expression, and Identify Novel Anti-tumor Agents

En kvantitativ analyse for å studere Protein: DNA interaksjoner, Discover Transcriptional regulatorer Gene Expression, og identifisere nye anti-tumor Agenter

DOI:
10.3791/50512-v

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06:43 min

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August 31, 2013

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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

Summary

Automatic Translation

Automatic Translation

Vi har utviklet en kvantitativ DNA-bindende ELISA-basert assay for å måle transkripsjonsfaktor interaksjon med DNA. Høy spesifisitet for RUNX2 protein ble oppnådd med en konsensus-DNA-gjenkjenning oligonukleotid og spesifikt monoklonalt antistoff. Kolorimetrisk deteksjon med en enzym-koblet antistoff substrat reaksjonen ble overvåket i sann tid.

Tags

Protein:DNA InteractionsTranscriptional RegulatorsGene ExpressionAnti-tumor AgentsDNA-binding AssayEMSAChIPD-ELISARUNX2Protein-DNA InteractionQuantitative AssayDrug Screening

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