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293 and MTT assay - (Oct/16/2001 )

Hi, I would like to know any protocol to assay MTT with HEK293T cells. I´ve proved, but formazan crystals don´t affix to the plate. Does anyone can help me?

-Sara-

do you use 24 well plates or 96 well plates? Try bio-coated plates. Also try to suck off the media / mtt solution with mediaa pasteur pipette attached to a rubber bob..this usually worksand the crystals should be fine...

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I am using MTS, which produces a soluble formazan product. The assay is dead easy, just add cells, then add MTS solution (from Promega), incubate, and read.
As I undersatnd, the MTS does not enter the cells. It is reduced on the outside of the plasma membrane. One problem that I have come up against is that it is linear w.r.t. viable cell number only over a narrow range.

-Stephen Pratt-