Hello everyone, Please I am trying to prepare different concentration of doxorubicin for my cancer cells line, Can someone help me with the calculations so I can continue my experiment. Thanks in anticipation !
Hello Innocent Agida
You dilute the doxorubicin stock 1:100 to obtain diluted stock of (1mM).
Use the formula C1V1=C2V2
C1= concentration of diluted stock (1mM)
V1= volume of diluted stock to be taken (X)
C2= concentration of working solution (10uM)
V2= Required final volume (say 1ml)
1000 uM x X = 10uM x 1ml
X = 1 x 10/ 1000 = 0.01ml = 10ul
So, for 10uM, you will add 10ul of diluted stock (1mM) to 990ul of culture media to obtain 10uM concentration in 1ml.
Similarly,
For 5uM, you will add 5ul of diluted stock (1mM) to 995ul of culture media to obtain 5uM concentration in 1ml.
For 2.5uM, you will add 2.5ul of diluted stock (1mM) to 997.5ul of culture media to obtain 2.5uM concentration in 1ml.
For 2uM, you will add 2.0ul of diluted stock (1mM) to 998ul of culture media to obtain 2.0uM concentration in 1ml.
For 1.5uM, you will add 1.5ul of diluted stock (1mM) to 998.5ul of culture media to obtain 1.5uM concentration in 1ml.
For 1.0uM, you will add 1.0ul of diluted stock (1mM) to 999ul of culture media to obtain 1.0uM concentration in 1ml.
However, for 0.5uM and 0.1uM concentrations of doxorubicin, the volume of diluted stock to be pipetted is very low and pipetting such small volumes will lead to error. So, you dilute the diluted stock (1mM) by 1:10 to obtain the drug concentration of 100uM.
So, for 0.5uM, you will add 5ul of 100uM solution of doxorubicin to 995ul of culture media to obtain 0.5uM concentration in 1ml.
For 0.1uM, you will add 1.0ul of 100uM solution of doxorubicin to 999ul of culture media to obtain 0.1uM concentration in 1ml.
Good Luck!
Ok @ S.Soman. Thanks for the response, making the 10 times concentration how do I do this? What volume of medium will require ×10 dilution with medium
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