proliferative responses

Joanne Thomas thomas at
Wed Sep 20 09:11:46 EST 2000

You can try MTT, a water soluble tetrazolium salt, which is a yellow
solution that is converted to an insoluble purple formazan when cleaved by
dehydrogenase enzymes. The insoluble formazan can be solubilized isopropanol
and then measured on a spectrophotometer. Sigma sells MTT (Product # M5655)
relatively cheap. In my hands it worked really well for mitogens but was a
bit less sensitive for recall antigens (we used it at 5mg/ml). Some have
used CD69 and flow for this also, correlations were good, however, you'll
have to deal with the issue that cellular activation doesn't always lead to
cellular proliferation.
-----Original Message-----
From: Jose Benito <jbenito1 at>
To: cyto-inbox
Date: Tuesday, September 19, 2000 6:57 PM
Subject: proliferative responses

>Hi everyone, I am trying to set up a colorimetric assay to measure
>lymphoproliferative responses to mitogens and recall antigens. I have tried
>the Alamar Blue without any sucess. I will probably try also the celltiter
>assay from promega. Does anyone have experience with this kind of assays?
>Can they really substitute the 3Hthymidine assay?
>Jose Miguel
>Hospital Carlos III
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