I have been asked to make 100 micromolar and added to the cell culture
I used this calculation : 1. Moles = Molarity (M) x Volume (L)
I decided to make 0.5 M as a stock solution this dilute it
moles = 0.5 x 0.001 L ( 1 ml) = 0.0005 moles
then,
Mass (g) = Moles x Molar mass (g/mol)
mass= 0.0005 x 430 = 0.215 g
so I will weigh 0.215 g and dissolve it in 1 ml of water.
so the stock concentration is 0.5 M I will dilute it to 100 micromolar
I will use M1xV1 = M2xV2
M1 = 0.5 M
v1= ?? unknown how much I will take
M2 = 100 mM
v2= 10 ml
by doing the calculation I will need to take 0.0002 ml = 0.2 microliter which is too low
my question: I will add 0.2 microliters to the cell culture plate that has 2 ml of media I am afriad that the drug will be dulit more because of the media and the concetration will be less than 100 mM
what your recommadation?
Also are my calcualtions correct?
Hello Meme Ali
I have not understood your question quite well. Nevertheless, I will try answering your question assuming that you need 100mM as your final concentration in 2ml of culture media.
You have decided to make 0.5M as the stock solution. I am assuming that the molecular weight of the drug is 430g/mol.
You may do as follows.
430g – 1M – 1L
215g – 0.5M- 1L
0.215g – 0.5M- 1ml
215mg – 0.5M – 1ml
Weigh 215mg of drug and dissolve in 1ml water to give 0.5M concentration.
Use the formula
M1xV1 = M2xV2
M1 = 0.5 M (500mM)
v1= ?? unknown how much I will take (X)
M2 = 100 mM
v2= 2 ml (final volume in the culture plate) (Not 10ml)
500mM x X = 100mM x 2ml
X = 0.4ml
So, you will not add 0.2ul to the cell culture plate that has 2 ml of media. Instead, you will add 0.4ml (400ul) of stock solution (500mM) to 1.6ml of culture media to make a total volume of 2ml of culture media, which you will add to the culture plate. This will give you 100mM of the drug concentration to be finally present in 2ml of culture media.
Best.
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