New to MTT.... help calculate - (Sep/06/2012 )
As I am new to MTT and a foreigner scholar in China, have to really strive the language barrier over here.... so am writting down my question here, I do know the protocol for MTT but could not find any help form internet that how to further calculate/interpret my readings for MTT I got on excel.
Can somebody please guide me that after getting the data (absorbance reading) on microsoft excel how can I actually calculate the cell viability rate. I mean what will be the formula applied or what i have to do....no nothing ....
Assay was performed in six replicates for blank, control and each concentration of drug.
I used a blank (no cell only mtt reagent+DMSO), control (cells+mtt reagent+DMSO), and various concentration of my drug in micromole (from 1um to 100um). Now I don't know what to do further with the absorbance obtained. I shall be thankful to the one who can teach me step by step calculations for mtt on excel to get the final results (i.e percentage survival).
R u an Indian? Coz I am an Indian also doing my Ph.D on drug targetting in breast cancer cell line MCF-7 and MDA-MB 231. Here are the things that might help u:
<*>First u calculate all the average of OD taken in all ur replicates.
<*>Now u note down in ur note book against respective dtug concentrations.
<*>Take the average of the CELLS+DMSO (0 micro mole or positive control).
<*>Now calculate the percentage of viability of each drug concentration against the average of positive control where it is assumed that the viability in control is 100%.
<*>Now u can plot the percentages and the respective drug concentrations in Excel and can obtain the graph.
Now u do these. If u face any prob then feel free to ask.
I would like to know if these applies to the MTS as well ??
I have cells+MTS reagent with Media is this my + control ??
i just posted a question here, read this post after that.
i wanted to know when is the ideaal time to add drug to the cells?
same day as u culture the cells or 24hrs later or should i wait for the cells to be concurrent.?