I am going to see the IC50 value of a powdered natural supplement. To treat the cells in 96 well plate, I have to dissole them in these concentration: 0, 0.25, 0.5, 1.0, 2.0, 4.0, 6.0, 8.0 and 10.0 mg/ml. I will plate the cell suspension in 10% serum containing RPMI growth medium. Should I also treat the cells with the natural supplement dissolved in the same growth medium? Any protocol with reference will be appreciated.
Hello Anwar,
You can evaluate IC50 by using MTT assay or any other techniques you might have used before.
But, In order to conduct such experiment for estimation of IC50 value, you should maintain the uniform environment i.e. growth medium for the cells to grow.
So, I will suggest you to prepare all of your dilutions in the culture medium only prior to start treatment. Also calculate the % of diluent (in which the powder was initially dissolved) which will be present on each dilution preparation. For control you must prepare media containg same dilution of solvent( in which the powder will be initially resuspended) that will be present in your test material containing medium. If water will be your solvent then culture cells in regular growth medium and treat as an internal control to calculate % toxicity and IC50.
Jaykumar Kambli Hi, thanks for your response. Can you please recommend me any of the solvent which can be used to dissolve any kind of powdered natural supplementary product?
Md Rafi Anwar Hello, Water will be the best solvent for any kind of study.
But, you should check the solubility of your compound and dissolve accordingly to prepare a stock. Some time Ethanol or DMSO can be used depending on solubility of respective components.
The ultimate thought is that solvent must be least toxic to cells.
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