non-radioactive cell growth assay

Rosemarie Ganassin RCGANASS at BIOLOGY.Watstar.UWaterloo.CA
Mon Mar 20 15:12:19 EST 1995

michael.micksche 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
> >Department of Genetics, SHM I-166  ecg at (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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