I check the viability of breast cancer cell lines(MDA-MB-231 and MCF7) with 10-OH-Camptothecin (CPT) to get the effective concentration of this drug.
I used RPMI 1640 for cell culture (our lab did it for a long time) and seeding to 96 well plate (3,000 cell/well) for 24 and 48-hr. After that, I use 10-OH-CPT for several concentration (1nM~10μM) in 200uL and incubate it for 24 and 48- hr. (10-OH-CPT dissolved in DMSO 1 mg /ml store at -70 degree, and dilute in media before use)
Final, 20 µL of MTT (5 mg/mL in PBS) was added to each well and incubated in a dark place at 37˚C for 3 hours. There after evacuating the contents of the plate and 100 μl of DMSO was added to each well and the plates were incubated for 20 min in a dark place at 37˚C. The absorbance was measured by an Elisa reader at 570–630nm wavelength. As a result, I get different results and I do'nt get a definite IC50! Especially in the MCF7 cell line.
Can anyone help me to solve the problem?
Thanks
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