IUBio

Replacement of "old" 3H-thymidine assay.

Kevin Law Law_Kevin_L at lilly.com
Thu Mar 25 08:58:52 EST 1999


In article <01be7608$679fe2a0$0c2b899b at jrh-05236.leo.dk>, "Jens Rainer
Hansen" <jens.rainer at leo.dk> wrote:

>I want to replace my 3H-thymidine assay on human lymphocytes, since it is
>to hard to perform when having a lot of samples.
>
>I have tryed alamarblue but it doesnt work with lymphocytes.
>The MTT assay should also be insensitive with these cells.

We have used CellTiter96 AQueous One Solution Cell Proliferation Assay
from Promega (www.promega.com) for both human and murine leukemia cell
lines for many years. It's an MTT like reagent which requires almost no
manipulation and you get good color development in 1.5-2 h. I couldn't
live without it - makes my life much easier. Might check it out. Should
have technical details available on their website.

>
>I have also tryed Brdu incorporation it seems ok, some times, but since the
>lymphocytes are non adherent it is nessesary to dry the cells on the
>microtiterplates with an hairdryer, and this procedure was not stable for
>me.
>
>Do you have experience with an easy assay to meassure DNA synthesis/cell
>proliferation on human lymphocytes (non adherent) using other methods than
>3H-thymidine, or methods where it is easy to handle the samples using
>3H-thyminine, that means not punching filters into vials etc.
>
>
>Yours Jens R. hansen

-- 
Kevin Law
kll at lilly.com
Eli Lilly and Company



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