Why TNF in dose of 100ng/ml doesn't kill my cells in MTT assay? and S1P doesn't protect these cells from necrosis?
The decision to undergo necrosis depends on the level of ATP. Your cells probably need a combination with TNFa to induce apoptosis. A simple way to check this is using cycloheximide with TNFa. 100ng/ml TNFa is too much. You may reduce it down to 1ng/ml to 17ng/ml.
MTT is an assay to evaluate viability, not cell death. Moreover, the dynamic range of MTT (and MTS) is very short, this means that you will not see small difference between groups. Maybe, you can use an other assay to evaluate cell death, like trypan blue exclusion assay, cell cycle (flow cytometry), LDH assay, etc.
MTT assay is suitable for evaluating viability or proliferation. If you want to measure apoptosis or cell death Annexin V staining is a good option. In any case you have to perform dose response curve to determine the optimal dose that result in cell death under your experimental condition.
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