I am doing MTT assay for my plant extract. after cell seeding how should i dilute my plant extract for better result. should i dilute it in that solvent in which it has been prepared or in cell culture medium.
Can i add my plant extract to the seeded cells directly or it should be added alongwith media in that it has been diluted.
Kindly suggest me .
Hi,
Regarding your first question of the dilution, I would strongly recommend you not to dilute your extract rather I would suggest you to use a 5 different volumes (in uL range). Doing this will be helpful for you to calculate IC50 value.
Regarding your second question, I would suggest you to perform the MTT assay separately for the solvent you are using for your plant extract. Later you can subtract the solvent's value from the actual reading and you can estimate the exact cytotoxic behaviour of you plant extract. Later, you can plot cell viability curve to calculate IC50 value.
Thank You Shivendu. Can you tell me one more thing that if i will dilute the plant extract in solvent rather than diluting in culture media , it will not starve the cells.
After cell seeding can we add our test extract to the cells without adding media. if we have to give 24hr incubation then in that case because of the absence of media it will not cause cell starvation?
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