I am curious whether MTT can be use for HTS or not? i need to determine the cytotoxicity of 50 different small compounds against various panel of cancer cell line (7 cancer cell line). Is it feasible to use MTT as HTS? IS there any other cytotoxicity assay to replace MTT and more reliable one.
I would highly appreciate if someone share some related information.
Hello,
Yes you can. In our lab we using MTT assay. Moreover in some case we using NR (neutral red). This assay give good results too.
http://www.sciencedirect.com/science/article/pii/S0378427405001967
definitely one can use MTT. But I would go for single step addition and read reagent like MTS or CCK8
You could - but MTT is for one thing carcinogenic (not so good for handling on a large scale) and multi-step so it won't be all that high throughput - but "high" is different to different people! If you want to stick with this, MTT has been replaced by MTS - still colorimetric but at least you don't have to add the solubilisation buffer and you can continuously monitor the development of the assay if you wish - for a short time anyway, which is helpful during assay development.
Coming from a pretty high HTS background myself, I'd suggest a look at Promega's CellTiter Blue (= Resazurin/resorufin) which can be more sensitive (can be read colorimetrically or as a fluorescent output) and most importantly for true HTS, a one-step addition assay. This way, you'll save cells (= less handling, maintenance reagents) and time.
Good luck!
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