mononuclear cells medium - I am stuck with the concentration of 2-mercaptoethanol (Jun/14/2010 )

hi all,
I am new in cell cultures and need to grow mononuclear cells produced from bovine blood
all that's written in my protocols is to include 50 micromolars of 2-mercaptoethanol in the medium.
how do I convert this into the amount I have to put is my medium (in microliters)
hope someone can help because the accuracy is important.
Gitit

Hi,

I have just made up a sterilised solution of 200 mM (0.2 M) beta mercaptoethanol in the past and added that to my culture media in a sterile biological safety cabinet.

Use the equation of Concentration 1 multiplied by Volume 1 = Concentration 2 multiplied by Volume 2

Say you will have the standard 500 ml of culture media. You know the final concentration of mercaptoethanol is 50 uM and the starting concentration is 200 mM. 1000uM = 1 mM. Therefore 200 multipled by 1000 = 200,000uM.

So:

200,000 uM x V1 (in ml) = 50 uM x 500 ml

V1 = 25,000/200,000

V1 = 125 ul of mercaptoethanol