I am doing the enzyme assay of my compounds for DPP-IV inhibition. My compounds are soluble in DMSO but not in tris buffer pH 7.5. When I make a stock solution (1mg/ml) in DMSO it is soluble but when I dilute it with buffer (for further dilutions) it gets ppted. I used vertex mixture to increase the solubility but it did not work with my compounds.
When I use the prepared solutions (in DMSO) for enzyme assay it again get ppted. According to my protocol I first add serum (10µl) then 40µl inhibitor solution and then DPP-IV substrate (made in tris buffer) 150µl.
Can I premix inhibitor solution and DPP-IV substrate and then add to micro-plate (by this way the compounds not ppted)? Please suggest some way to carry out my enzyme assay with these compounds.
Please suggest some way to dissolve my compounds.
Regards
I had once a similar problem and what I did was making a first dilution in water and the a second dilution in the buffer. It is not the opltimal procedure, but at least in that particular case it helped me to solve my solubility problem. Using Triton-X100 might also help, but first you should check if that does not affect the activity of your enzyme.
I had the same problem and I solved it by using Triton X-100 in a 0.5 %. I hope it will work for you.
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