What is the impact of DMSO on calcium influx, and why does it lead to a significant decrease in fluorescence when compared to the control group without treatment during ATP-induced Fluo-4 assay results? Results attached. What alternative solvent can be used to dissolve a drug without causing interference with calcium dyes
DMSO can significantly decrease fluorescence intensity in Fluo-4 based calcium assays. This is likely due to:
- DMSO is hygroscopic and absorbs water from the assay buffer, which can reduce Fluo-4 fluorescence intensity.
- DMSO can directly interact with Fluo-4 and cause fluorescence quenching through various mechanisms.
- High concentrations of DMSO can disrupt cell membranes and calcium channels, reducing the influx of calcium upon ATP stimulation.
- DMSO is amphiphilic and can bind to hydrophobic pockets on proteins, altering their confirmation and function.
To avoid these interferences, an alternative solvent such as ethanol or methanol can be used to dissolve compounds for calcium assays.
And appropriate vehicle controls should also be included.
Thanks Salah.
I used only 1% DMSO. May be i will try other solvent to dissolve my drugs
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