I have been testing some inhibitors against CT26 cancer cell lines using Resazurin cell viability assay! I observe that higher inhibitor concentration showed % cell viability higher than slightly lower inhibitor concentration!
For example: at 2 microM % cell viability is 20 %
at 0.2 microM % cell viability is 5 %
Does higher inhibitor concentrations stimulate the cell viability?
Thanks in advance for your answers
If its inhibitor reacts with resazurin, this would explain the reason for the higher fluorescence with the higher doses of the inhibitor. Ideally you would perform (as a control) a reaction of the inhibitor you wish to test in PBS (for example) add Resazurin and observe if this produces any fluorescence. Another option would be to use MTT as a method to measure cell viability.
Solvents used for dissolving inhibitors affect the colorimetric asays. for eg. the vehicle DMSO should be maintained at 0.1 % to avoid with assay as well as pathway interference. At times RPMI media used in cell cultures too can affect the viability assays.Check the concentration of the solvent used. As well as read the pamphlet for assay kits to know the interference lists.
depending on the nature of your inhibitor (competitive or not, reversible or not) the answer is:
maybe / maybe not.
- Badges
- Science topic