how to dissolve compound in MTT assay???

[gajera888]

hello frends...

i am checking cytotoxicity of some compounds using MTT assay.

my compound is soluble in Dichloromethane and DMSO. but when i add it in RPMI 1640 media in 96 well plate...it precipited out. and i got the wrong results. may be my compound is in precipited form during incubation time period of 72 hrs.

wht shud i do next????

[luisillo]

I've seen some reagents that are only soluble in DMSO (in my case PMA). The problem in MTT assay with DMSO (I don't know about dichloromethane) is that DMSO per se is toxic for the cells.

What you have to do in these cases is to add a similar volume of DMSO to dose 0 (untreated control), so that cytotoxic effects can be correctly attributed to your compound and not to the solvent.

In my case, the label of PMA said that you should initially dissolve the lyophilized PMA in DMSO and later you could take some to prepare aqueous solutions in media. I don't know if your compound may work in the same way, but I recommend you try it, always remembering the above stated thing about the untreated control.

I hope you can solve your problem. The best of luck.