We were performing Glutathione-S -transferase assay using 1-Chloro-2,4-Dinitrobenzene as a substrate. However, it was practically precipitating while preparing a 30mM solution. We dissolved fin methanol at first and made it up with distilled water, which actually resulted in precipitation
Dear Tincy,
thank you very much for sharing this interesting technical problem with the RG community. My general suggestion would be to strictly follow a published protocol for the glutathione-S-transferase assay. 1-Chloro-2,4-dinitrobenzene is insoluble in water, and there is no way to circumvent this. The compound is soluble in methanol, ethanol, diethyl ether, and acetone. Just add only as much water that no precipitation occurs. Slight warming or ultrasonication might help, but in the end you want to have a clear solution which is stable at room temperature. 1-Chloro-2,4-dinitrobenzene will remain insoluble in water, no matter what we do. Thus look for an established protocol in which the preparation of a solution in methanol / water is described in detail.
Good luck with your experiments and best wishes, Frank Edelmann
Frank T. Edelmann We tried the addition of ethanol and water alternatively and slowly and stopped just at the point of precipitation. The total volume was made up of both ethanol and water without precipitation, exploring the maximum water solubility. Thank you very much for your suggestion
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