I'm working on a project testing a drug on 4 different cell lines ( RAJI- SW626-BT20 and PC3). my drug has shown a cytotoxic effect on all the four types using MTS antiproliferation assay. I'm thinking to do another experiment like western blot that could help in providing more evidence to confirm that my drug is actually affecting the cancer cell cycle an some point by causing arrest or apotosis.
It is rather cytostatic effect. To check if it causes cytotoxic effect, you may either do: double staining with Hoechst/PI, annexin assay, caspaze 3/7 assay or check the cleavage of PARP on western blot. Then it would be good to check p21,p16 on WB (cell cycle arrest) and cell cycle analysis on flow cytometry. SubG1 should also show the proapoptotic effect of the drug.
hi,
for testing cytotoxic effect you can apply MTT or Neutral red essays, also you may conduct Ethidium bromide/acridin orange or annexin-V-flous/PI staining tests for apoptosis study, followed by flow cytometery.
For more information related to your work visit this article:
Article Brevinin-2R and Derivatives as Potential Anticancer Peptides...
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