I do MTT assay for cell viability. I used MTT solution 1mg/ml, 2h incubating and then treated DMSO to formazan. I read 595nm.
In march, O.D was fine ( repeated 6 times ). So i perform experiments. The recent, i perfomed FACs, have never seen effects Annexin V. It is strange, so i do MTT assay this week but that wasn't affect too.
Cell is A375 carcinoma, material stock disolving DMSO.
Cell passagee13. i think cell problem because the result extremely changed. But any other problems?
I can not speak English very well... i'm so sorry
Hi Jaehan Lee
The error might be seeding error. Hence, ensure the
equal numbers of cells in all the well and further, at the end of treatment, check the morphological changes in treated cells as well as in control control under microscopy.
Thanks Prasath Manogaran
I chenked each weel after seeding, but i saw almost same amount.
But i think seedings doesn't effect MTT result like my experiments.
Then, After treatment check the cells and do Annexin V staining. if there is no cell death, check the concentration of your drug and optimize the cell density.
All the best
Prasath Manogaran
Ok i will check death percentage before Annexin staining.
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