Rapamycin Concentration for Cell Culture - (Jan/22/2014 )

Hello guys,

I am an undergrad student and I have been working to induce autophagy in C2C12 cell lines but it has been 3 months without any success 

My cells are passage number 930129383912 so it could be that the cells are bad but I have started to doubt whatever I am doing. 

It has been shown that 250 nM concentration of rapamycin induces autophagy in my cell line.

(if you have time) could you please calculate the dilution that I need to see if I got it right?

The stock that I have is 1mg/ml dissolved in DMSO. 

mw of rapamycin is 914.2 g/mol 

I am doing my experiment in 96 well plate and I put 100 microleters in each well. My rapamycin HAS to be dissolved in 1 microleter of DMSO.

By that I mean, I add 1% DMSO (rapamycin disolved) media to each well. 

(what I am doing is that I dilute my sample ~43 times and I add 10 microleter in 1 ml media) then put 100 microleter of that solution in each well.

Thank you all and any help would be appreciated because I am very frustrated 

Hossein

So - you have 1 mg/ml rapamycin which you add to medium to get a 1% solution (of the solute, note that this might be too much for most cell lines.  DMSO often has effects down at the 0.1% range)?

Anyway - 1 mg/ml * 0.01 = 0.01mg/ml or 0.01 g/l ... can you take it from there?

c=n/v, n= mass/molar mass...