Hi all,
I'm doing a bunch of Cell Titre Glo assays to assess the effects of several inhibitors on 2 cell lines. Problem is, for some of the wells they seem to be luminescing less than the background control (media only) wells, some of the time. So I end up with negative normalised values. What would cause this? It's not consistent between replicates or repeats.
Thanks!
First of all, non-uniform luminescence may be caused by uneven cell seeding, temperature gradient, or edge effect. If you are confident about these conditions, you can check for contamination in your media. It might be the fact that the inhibitor is also killing the microbes along with your cells in the test wells but not in media only cells. Secondly, check the shelf-life of your assay kit to figure out any problem with the kit. All the best.
Thank you Nabodita. I hadn't thought about media contamination. The kit is pretty new so I'm hopeful it's not that. I'll investigate further!
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