Novel and Efficient Protocol for DNA Coating- Based Identification of DNA-Protein Interaction by Antibody-Mediated Immunodetection

سال انتشار: 1399
محل انتشار: مجله گزارش های بیوشیمی و زیست شناسی مولکولی، دوره: 9، شماره: 3
کد COI اختصاصی: JR_RBMB-9-3_003
زبان مقاله: انگلیسی
تعداد مشاهده: 202

نویسندگان

Regeneration and Stem Cell Biology Lab, Centre for Molecular and Nanomedical Sciences, International Research Centre, Sathyabama Institute of Science and Technology, Chennai, Tamilnadu, India.

Johnson Retnaraj Samuel Selvan

Regeneration and Stem Cell Biology Lab, Centre for Molecular and Nanomedical Sciences, International Research Centre, Sathyabama Institute of Science and Technology, Chennai, Tamilnadu, India.

Saravanakumar Venkatachalam

Regeneration and Stem Cell Biology Lab, Centre for Molecular and Nanomedical Sciences, International Research Centre, Sathyabama Institute of Science and Technology, Chennai, Tamilnadu, India.

Jackson Durairaj Selvan Christyraj

Regeneration and Stem Cell Biology Lab, Centre for Molecular and Nanomedical Sciences, International Research Centre, Sathyabama Institute of Science and Technology, Chennai, Tamilnadu, India.

چکیده

Background: Studying protein-protein and protein-DNA interactions are prerequisites for the identification of function and mechanistic role of various proteins in the cell. Protocols for analyzing DNA-based Protein-Protein and Protein-DNA interactions are complicated and need to be simplified for efficient tracking of binding capabilities of various proteins to specific DNA molecules. Here, we demonstrated a simple yet efficient protocol for the identification of DNA coating-based Protein-DNA interaction using antibody-mediated immunodetection. Methods: Briefly, we have coated specific DNA in the microtiter plate followed by incubating with protein lysate. Specific protein-DNA and/or protein-protein bind with DNA interactions are identified using specific fluorophore-conjugated antibodies. Antibodies are used to detect a protein that is bound to the DNA. Results: Fluorescent-based detection identifies the specific interaction between Protein-DNA with respect to coated DNA fragments. The protocol uses indirect conjugated antibodies and hence the technique is sensitive for effective identification of Protein-DNA interactions. Conclusions: Based on the results we conclude that the demonstrated protocol is simple, efficient and sensitive for identification of Protein-DNA interactions.

کلیدواژه ها

Keywords: DNA coating, Lamin A, Protein-DNA interaction.A, trf2

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