I am trying to dissolve SGZ to be used as substrate for Laccase activity assay. I tried dissolving SGZ in absolute ethanol but it is not dissolving completely, more ever it is forming suspension in the assay mixture creating fluctuations in absorbance.
Please suggest me technique to dissolve SGZ completely.
Syringaldazine, 0.56 mM stock solution: Rinse a 50-ml
volumetric flask with water and ethanol to remove any soapy
residues. Weigh 10.0 mg of syringaldazine in a weighing boat
and transfer to the volumetric flask. Add 96% ethanol to the
mark and stir until the syringaldazine is dissolved
(approximately 3 h). The solution must be stored in a dark
bottle in a refrigerator.
Syringaldazine, 0.22 mM working solution: Rinse a 10-ml
volumetric flask with water and ethanol to remove any soapy
residues. Transfer 4.0 ml of syringaldazine stock solution to the
flask and add water to volume. The solution can be kept in a
dark bottle for up to two hours at room temperature
To prepare a 1 mM solution, you could dissolve the reagent in 95• ethanol, under stirring. Gentle warming of the flask will speed the dissolution process. The obtained solution will remain stable for a very long time when stored in the fridge, without any precipitation.
I suppose sonication will assist in dissolving SAZ in absolute ethanol. Give it a try using a sonication bath. Good luck!
SGZ is a good substrate for laccase enzyme but the problem it's to dissolve it well!
I propose for you two types of solution:
DMSO: SGZ is highly soluble in this solution and it could be dissolved immediately in this fraction. But if you use some buffer such phosphate buffer, you will have precipitation problem.
Absolute ethanol: SGZ is soluble in ethanol but it need long time for dissolution, you can help with sonication or vortex. The problem here that the ethanol is highly evaporable and the concentration could change during the reuse of the substrate! Then it could be better to dilute the SGZ-ethanol solution by water to stabilize a little bit the solution and inhibit the evaporation.
DMSO though a good solvent it also quenches the laccase generated radicals - if incubated for a long time- all depends on purpose
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