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Interpretation on absorbance reading from 96 well MTT assay

MTT 96 well absorbance assay

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#1 Frazermb

Frazermb

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Posted 23 October 2013 - 06:56 AM

Hi,

 

I have just carried out an MTT assay using THP-1 cells.

 

I added 10ul of MTT to 200ul of cells in media and incubated for 90 minutes.

 

After incubation the solution was removed an 100ul of DMSO was added and the plate was then read at 570 in the spec.

 

using Beer Lamberts Law i had to work out the concentration of Formazan.

 

c=A/(ExL)

 

c= 0.345/ (13x0.696)

 

However the resulting answer was 0.039.

 

The concentration is supposed to be in uM.

 

What step am I missing with regards to the equation.

 

Thanks

 

Frazer







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