I want to detect that my proliferating cells underwent a DNA Damage after treatment with drugs. I already know the H2AX method but looking for other methods. Many thanks in advance!
I hope the paper attached will help you more.
Tunnel assay, comet assay, H2AX phosphorylation, immunocytochemistry and few other methods are mentioned in these.
Detection of DNA damages can be done after staining with propidium iodide with a flow cytometer or a fluorescence microscope. Alternatively, PCR and agarose gel electrophoresis can be done. You can check the following publications for more information: Article Strategies for the evaluation of DNA damage and repair mecha...
Chapter Methods for the Detection of DNA Damage
Good luck - CL
I hope the paper attached will help you more.
Tunnel assay, comet assay, H2AX phosphorylation, immunocytochemistry and few other methods are mentioned in these.
Hi Umran, The easiest method you can use is DNA fragmentation assay. See:
Article An overview of apoptosis assays detecting DNA fragmentation
https://www.creative-bioarray.com/support/dna-laddering-assay.htm
You can also use DAPI staining to see nuclear fragmentation, which occurs at advanced stages of apoptosis. One standard protocol is here: https://www.genscript.com/tech_guide/TM0358.pdf
Thanks
Arun
You can do too a cytockinesis micronucleus assay or an in vivo assay.
OECD (2016), Test No. 487: In Vitro Mammalian Cell Micronucleus Test, OECD Guidelines for the Testing of Chemicals, Section 4, OECD Publishing, Paris, https://doi.org/10.1787/9789264264861-en.
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OECD (2016), Test No. 474: Mammalian Erythrocyte Micronucleus Test, OECD Guidelines for the Testing of Chemicals, Section 4, OECD Publishing, Paris, https://doi.org/10.1787/9789264264762-en.
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You can detect the fragmentation of DNA by staining your cells with PI and analyze using flow cytometer
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