non-radioactive cell growth assay
RCGANASS at BIOLOGY.Watstar.UWaterloo.CA
Mon Mar 20 15:12:19 EST 1995
michael.micksche at univie.ac.at (Thomas Mohr) wrote:
> In article <3ka4pk$52r at usenet.INS.CWRU.Edu>, ecg at po.CWRU.Edu (Edward C. Goodwin) says:
> > If anyone has any suggestions for non-radioactive cell growth assays I would
> >greatly appreciate it if you would drop me a line. Our lab has been studying growth
> >arrest upon expresssion of certain viral proteins and uses tritium incorporation as an
> >assay for DNA synthesis. I seem to recall an add for some ELISA assay that would measure the
> >same thing, so if anyone out there in cyberland has any experience with this, positive or
> >negative, I'ld really like to hear about it.
> > Thanks in Advance,
> > Ed Goodwin
> >Edward C. Goodwin PhD ecg at po.cwru.edu
> >Department of Genetics, SHM I-166 ecg at po.cwru.edu@cunynm (bitnet)
> >Yale University School of Medicine "Facts do not cease to exist
> >333 Cedar Street because they are ignored."
> A very good and accurate MTT-Assay Kit is available from Biomedica, Vienna, Austria.
> The procedure is quick and simple, it encompasses only dissolving the substrate in a buffer,
> pipetting into the wells of 96-plates, incubation (approx 2-3hrs) and measurement of the OD 450.
> The regressioncurve between cellnumber/well and OD 450 is almost linear from 1000 to 5000 cells/well
> (Test line: SK-MEL 28) and the coefficient of standarddeviation is approx. 5-10% .
We've used another assay for measuring cell proliferation.
It's simple, but requires access to a microplate fluorometer,
such as the CytoFluor 2350. Bisbenzimadazole, or Hoechst 33258,
allows measurement of the amount of DNA in each well.
See Journal of Tissue Culture Methods, 1994, 16: 133-142. We're using
fish cells but the assay will work with any cell type.
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