I am studying PBMC cells through CD3/CD28 co-stimulation, and adding Acetylcorynoline (10, 5, 2.5, 1.25) μM at the same time.
The reference is PMID: 31724152.
After the first treatment of PBMC cells with Acetylcorynoline, the absolute count of T cells was detected by flow cytometry, and the IC50 of Acetylcorynoline was calculated to be 5μM.
Then treated with Acetylcorynoline for the second time, and the results showed that the same Acetylcorynoline concentration did not achieve the same inhibitory effect. As shown in the screenshot.
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Lucian Miles Some variability between runs is expected, so I’d recommend performing the assay in triplicate. That way, you can calculate the mean and standard deviation to more reliably determine the IC50. Also, depending on your experimental design, it may be helpful to normalize your T cell counts—either to the vehicle control or total PBMCs—to account for differences in cell input or recovery. This might give a clearer picture of the Acetylcorynoline’s effect.
All the best!
Differences in cell response to drugs are often the result of many factors, rather than a single factor that can be controlled.
It is necessary to synchronize PBMC batches and culture systems as much as possible, record detailed operation procedures, and reduce fluctuations.
In particular, the storage conditions of Acetylcorynoline require protection from light. When the solution is stored at -80°C, please use it within 6 months, and when stored at -20°C, please use it within 1 month.
The answer to this question comes from MedChemExpress (MCE) Technical Support.
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