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determining optimal cell count for MTT

cell count MTT assay

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#1 Shrubal

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Posted 20 September 2011 - 11:39 PM

Hi!

I'm new to the MTT assay and from what I've read, I have to first determine the optimal cell count before performing the assay with my test sample. None of the protocols that I've read for this [inlcuding the ATCC protocol attached], ask you to add any cytotoxic compound before adding the MTT. How then will the formazan crystals be formed if I'm going to plate cells with >90% viability? Or am I missing out on something?

Thanks

#2 KevinK

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Posted 21 September 2011 - 07:23 AM

With the MTT assay, the viable cells convert the substrate into formazan crystals. Therefore, the toxic agent is not needed for establishing optial cell nubmer. You want to make sure you are within the linear range with your cells and timepoints. Please feel free to look over the manual for the Promega version for more information: http://www.promega.c...y Protocol.ashx

Also, please do not hesitate to contact Promega Technical Services with any questions you may have.

Kind Regards,
Kevin
Promega Corporation
Madison, WI

#3 Shrubal

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Posted 21 September 2011 - 09:14 AM

O right! I'm sorry my bad.. i didn't quite read the principle properly and therefore misunderstood how the assay works.. Thanks Kevin





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