Proliferation assay needed

Jay Vasudevan jv4s at avery.med.Virginia.EDU
Tue Oct 3 12:16:07 EST 1995


In article <01HVZ1O4L4FM8X5NO2 at MSdisk.WUstl.EDU>,
(To:  Nikolai) <troianovsk_s at MSDISK.WUSTL.EDU> wrote:
>>Date:    1-OCT-1995 03:13
>>From: saliou at garnet.berkeley.edu (Claude SALIOU)
>>Description: Proliferation assay needed
>The methods used different approaches. In my point of view thymidine
>incorporation is the only direct meassurement of prolifiration as cell`s
>growth.
.......
>MTT assay:
.....
>Thymidine uptake:
......
>Nikolai

if you have access to a fluorescent plate reader then the membrane
permeant DNA binding dye Hoechst 33258 can be used. It is much quicker
(freeze/thaw lysis=1h, incubate with dye, read). You will have to
standardise the fluor. unit with cell number at some point but once
that is in place, it should be pretty straight forward
It is not as sensitive in terms of raw numbers (for my cell types I
get an increase of about 600 AFU over 2 orders of magnitude of cells
(1000-100000)/well) as thymidine, but if you can use this technique I
think it is a lot easier.

YMMV
-- 
Jayanand Vasudevan					(804) 982-1891
Lab of Near Geniuses  .......I'm still looking for my invincible summer....
Dept. Path./Biochem., Box 214, UVa HSC Charlottesville VA 22908
http://www.med.virginia.edu/~jv4s



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