I want to induce blockade with DPCPX and SH58261 (Selective A1 and A2A adenosine receptor inhibitors, respectively) in cultured L6 rat skeletal muscle cells as a part of an inhibitory assay. Both inhibitors are insoluble in water and therefore, the solubility is given in DMSO (Dimethyl Sulfoxide). I just wanna know the best DMSO percentage (100% DMSO or a dilution such as 0.5% DMSO) efficient in preparing the inhibitory solutions in required concentration.
Hello Dilan, You could try making a 1000X stock in 100% DMSO. If you do so, then the final concentration of DMSO in your media will be 0.1%, so you can avoid/minimize any untoward effects from DMSO itself. You may also want to check any relevant literature to see what conditions others have used. Once you have prepared your 1000X stick, store it frozen in aliquots.
Bruce
You may want to do a solvent control in addition as well. Our more sensitive (stem-cell like progenitor) cells showed some stress response with 0,2% DMSO alone.
Thank you Dr. Bruce, Samantha and Ernst for your valuable comments and recommendations. I hope to go ahead with a final concentration of 0.1% DMSO while using the same concentration with the vehicle (KRPH, pH 7.4), but without inhibitors as the control.
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