Hi,
I'm going to do an experiment in which I need a final concentration of 3 mM of a probe in a 200 μL cell lysate.
If the probe has a molecular mass of, say, 74 g mol-1 I have derived that I need to add a final volume of
$$V_1\cdot C_1=V_2\cdot C_2\Rightarrow V_1=\frac{V_2\cdot C_2}{C_1}=\frac{V_2C_2MV}{m}$$
- as $$C=\frac{n}{V}=\frac{m}{MV}$$.
To attain a mass of my probe I can weight off I decide to dissolve 5 mg of it in 800 μL PBS (buffer), so I get
$$V_1=\frac{200\cdot 10^{-6}L\,3\cdot 10^{-3}mol\,L^{-1}74.085g\,mol^{-1}800\cdot 10^{-6}L}{5\cdot 10^{-3}g}=\sim 6 μL$$
To me this seems like a reasonable volume of my dissolved drug to add to the lysate, but I am uncertain if it will truly give me a final concentration of 3 mM. Maybe I've only complicated matters with the formula as I'm not sure I use it correctly - especially regarding the dilution. Thoughts?
I hope I've made myself somewhat clear
Thanks!