RE; Non-Radioactive Cell Proliferation Assay

tylee tylee at ITIS.COM
Tue Dec 9 20:01:30 EST 1997


Dear Itabajara da Silva Vaz Junior,

The CellTiter 96 AQueous assay works by utilizing reducing compounds (such
as NADH & NADPH) produced by living cells to convert a tetrazolium compound
into a colored formazan product. The colored product is detected at 490nm
using a standard 96 well ELISA plate reader. The assay will work for
detecting almost any type viable cells, including lymphocytes. You simply
add the mixture of reagents provided (i.e. a solution of MTS tetrazolium
and PES electron transfer reagent), incubate, and record absorbance that is
directly proportional to the number of viable cells. Because the assay
relies on the cell's metabolism to produce "color", (and lymphocytes
generally have a lower metabolic capacity/less cytoplasm) the assay
generally requires more lymphocytes to produce an equivalent amount of
490nm absorbance, compared to something like HeLa cells.

The chemical structures of the tetrazolium and formazan, downloadable
copies of technical literature, and several references that have reported
using this technology are available from Promega's web site
www.promega.com.

I have successfully used the tetrazolium technology to assay growth factors
for several years and have not found an easier system to use.

Ty Lee



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