My compounds are soluble in Acetone, DMSO. For invitro assay the compound solution is to be made in assay buffer supplied with the kit. But the compounds are not getting dissolved even after vortex. I am thinking of adding 1% DMSO, but this is not in procedure. It may interfere with the assay result.
Contact the Manufacturer of the Kits.
They will be having their own scientific team and they may have a solution for your problem.
VJ.
Dear Manoj,
In general you can use upto 5% DMSO without affecting the enzyme activity, but in case of PTP 1B one can go till 15 % without affecting the activity. For more details go to the link.
http://www.perkinelmer.co.jp/Portals/0/resource/products_ls/reader/pdf/LC3000-AP-204.pdf
I suggest you to start with 1% and increase upto 5% to dissolve your compounds.
Best regards,
Rajnish
I would prepare a stock solution of your compound in pure DMSO than diluting it with the buffer. (e.g. dissolve the small molecule in DMSO that goes easily probably to get a 1-20mM solution and you can use a few uL or more to get the desired concentration in buffer). And this method is not equal with the previously mentioned (adding DMSO to the buffer of interest and than try to dissolve. If you are interested I can elaborate it on if not just use the info). good luck
As mentioned above 1% DMSO should usually be sufficient to dissolve. You could also use a blank with 1% DMSO to compare the effect in case you have not yet done the experiment. Good luck
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