Reports of Biochemistry and Molecular Biology
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Basic Sciences
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10.61186/rbmb
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1399
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gregorian
2020
10
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Novel and Efficient Protocol for DNA Coating- Based Identification of DNA-Protein Interaction by Antibody-Mediated Immunodetection
زیست شناسی ملکولی
Molecular Biology
مقالات اصلی
Original Article
<div style="text-align: justify;"><strong><em>Background:</em></strong> 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.<br>
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<strong><em>Methods:</em></strong> 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.<br>
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<strong><em>Results:</em></strong> 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.<br>
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<strong><em>Conclusions:</em></strong> Based on the results we conclude that the demonstrated protocol is simple, efficient and sensitive for identification of Protein-DNA interactions.</div>
DNA coating, Lamin A, Protein-DNA interaction.A, trf2
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269
http://rbmb.net/browse.php?a_code=A-10-509-1&slc_lang=en&sid=1
Niranjan
Chellathurai Vasantha
100319475328460015387
100319475328460015387
No
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
johnnbt@sathyabama.ac.in
100319475328460015388
100319475328460015388
Yes
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
100319475328460015389
100319475328460015389
No
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
jacksondurairaj@sathyabama.ac.in
100319475328460015390
100319475328460015390
No
Regeneration and Stem Cell Biology Lab, Centre for Molecular and Nanomedical Sciences, International Research Centre, Sathyabama Institute of Science and Technology, Chennai, Tamilnadu, India.