I'm new to this assay, and I originally only plated 5000 cells/well, using 1% FBS and media with no phenol red. The abs values I receive are only around 0.250 at 490nm, is this acceptable when the assay solution+media control gives me an abs of 0.150? I might try plating at a higher cell density but I'd like to hear everyone's thoughts on this.
Have you tried running an LDH standard using positive and negative controls with different cell densities? The higher the density, the greater the abs values. For a negative (cells w/ just media) control, 0.250 seems fine. The positive control should be much higher and when running the standard, you will see a bigger difference between the positive and negative as the cell density increases, which gives a bigger range to view the toxicity of substances you are testing on the cells. We usually plate around 20k cells/well
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