Hi, every! I am testing the inhibition effect of anticancer drug by ＭTT assay, which my anticancer drug has strong reducibiity. principle of MTT assay is that Succinate dehydrogenase in Mitochondria reduce MTT to insoluble formazan.
The procedure of MTT has discarded the supernatant of the medium and drug, but the uptaken drug and drug attached on the surface of well and cell membrane will reduce MTT and produce a fake result. so how can I improve it? Or may I assume that the amount of attached and uptaken drug is small which could just ignored? It makes me headache.
I would be grateful If anyone could give me a suggestion on the issue.
Kind regards
Mingjie
I suggest you the XTT test .. the same principle but the detector is different.
That is, as you said, a problem. If your compound is water soluble use washing with cold PBS: 3 times for 5 min with gentile rocking. This will get rid of egzogenously present test chemical but it will still be present in the cell if it is able to enter it. You will not be able to remove it from the cells so if it enters the cells you will need to change your test to something not based on mitochondrial reduction.
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