Hi Fitria,

I assume that at this stage your purpose is only to see whether your extract is having any anti-cancer activity or not? You just wish to see if the cell line is being affected or not. If this is so, then you need not determine the exact concentration and may even use known volume of the extract. For example, you may start with say 10ul and go upto 100-200ul to begin with. You may also go by serial dilution if you wish to. You may just relate the toxicity with volume used.Once you find that your extract is having activity, then you may go for quantification of your active principle. But do remember do keep a methanol control as your extract is in methanol.

Alternatively, if its possible then you may nitrogen dry your crude extract and weigh the dry residue left. It may then be redissolved in the desired solvent to get a stock of known concentration which may then be used for your MTT studies.

Your question is very vague. If you could explain what you are starting with and where your extract is coming from, maybe I can help you. However, as of now, it is difficult to give you any suggestion. Do your extracts contain your cells and are you trying to determine the viability of your cells in the extracts? Or are you exposing the cells to the extracts?

i want to investigate anticancer activity methanol extract from Ascidian (marine invertebrate) against cell lines using MTT assay. i don't how to determine extract concentration for MTTassay

Here is a protocol that was published over the internet. You can follow this protocol since this uses cancer cell lines as well. If your tests do not yield a good result, make the necessary adjustments in the various parameters such as the number of cells and the concentration of your extract. Let me know if you need more help.

www.rsc.org/suppdata/dt/b9/b927129p/b927129p.pdf‎

thank you Mrs.Aguilar

Hi Fitria,

I assume that at this stage your purpose is only to see whether your extract is having any anti-cancer activity or not? You just wish to see if the cell line is being affected or not. If this is so, then you need not determine the exact concentration and may even use known volume of the extract. For example, you may start with say 10ul and go upto 100-200ul to begin with. You may also go by serial dilution if you wish to. You may just relate the toxicity with volume used.Once you find that your extract is having activity, then you may go for quantification of your active principle. But do remember do keep a methanol control as your extract is in methanol.

Alternatively, if its possible then you may nitrogen dry your crude extract and weigh the dry residue left. It may then be redissolved in the desired solvent to get a stock of known concentration which may then be used for your MTT studies.