Trypan Blue - Multiwell Apoptosis Assay? (Dec/12/2006 )

Hi

I am looking into setting up a apoptosis assay using trypan blue in a 96 well plate ie grow cells in the plate, add trypan blue, wash off excess dye and then measure "blueness" using a spectrophotometer. Does anyone know a protocol for such a assay or the wavelength that should be used for trypan blue??

any help is much appreciated

I used to read at 590 nm in plate reader.

What about using a caspase assay, the definitive assay for apoptosis.

Trypan blue can not distinguish between necrotic and apoptotic cells. It will not tell you whether the cell death is necrotic or apoptotic.

However if you want to find out absorbance maxima of soluble trypan blue, you can just run a spectrum of soluble trypan blue on spectrophotometer and find out the peak point.