I intend to test the activity of drug with LEUKEMIA, but I have not dealt with this kind cell line before
Conventionally, like MTT assay for adherent cell like HELA, we will wash and aspirate off the solution so many times before adding MTT solution.
For LEUKEUMIA, a kind of suspension cell, Can i use any method without washing away the cells???
Thank you for your attention.
After incubation, you can add MTT (5mg/mL) and incubate for 4 hours. Then you must centrifuge the plate at 800 rpm for 10 min and remove the supernatant. Then add a total of 100 µL of dimethylsulphoxide (DMSO) to each well to solubilize the formazan crystals. You can see this method in this article:
http://www.scielo.br/pdf/bjps/v53n1/2175-9790-bjps-53-01-e15210.pdf
Teol Wong Because of the limited equipment in laboratory, it had better if i can use some MTT steps available with some improvement for the Leukemia cells. Anyway, thank your your advice
Nasim Dana great! but i could not find out the plate centrifuge in my department. so is it possible if i transfer the cell to eppendorf then centrifuge it???
We use Alamar blue assay in my lab, it is some derivate of MTT assay, easy protocol. We seed out the cells with drugs/control in 200 µl and after 3 days, we add 20 µl Alamar blue solution - and after 4-8 h (depending on the cell type and metabolism activity) we measure at 570 and 600 nm without any centrifugation step before, just make sure that solution is equally distributed by shaking function of our ELISA reader.
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