Hi everyone, While performing a cell experiment, I noticed that precipitation occurs when I dilute the stock solution of Clozapine N-oxide (CNO, HY-17366) with PBS. This raises concerns about its solubility and stability in aqueous buffers. Has anyone else experienced this issue with CNO? Could the precipitation be due to poor solubility in PBS, pH differences, or an interaction with buffer components? What would be a better dilution method or solvent to keep it fully dissolved and bioactive for cell culture experiments (e.g., using DMSO or culture medium first)? Thanks in advance!
Hello Chloe Sun
You may use DMSO to make the stock solution of Clozapine N-oxide (CNO). CNO (freebase) is soluble in DMSO up to 100mM. Further dilutions may be made with culture media to maintain the final DMSO concentration in culture at ≤ 0.1% since DMSO is toxic to cells above 0.1%.
If you need to work with PBS, you should use the hydrochloride salt of CNO which is much more soluble in water. CNO hydrochloride has a solubility of approximately 1mg/ml in PBS.
Best,
Clozapine N-oxide has poor water solubility. Therefore, if you use PBS for dilution at higher concentrations, precipitation may occur. For cell-based experiments, it is recommended to directly dilute Clozapine N-oxide to the working concentration using culture medium, rather than preparing intermediate dilutions with PBS. Additionally, after diluting with culture medium, it is advisable to use ultrasonication to promote dissolution. Ensure that the solution is completely clear before replacing the medium and adding it to the culture plate.
The answer to this question comes from MedChemExpress (MCE) Technical Support.
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