I have one bottle containing 500ml F12K. In order to make a complete medium, I have to add in 0.1mg/ml heparin and 0.05mg/ml ECGS. How do I prepare heparin and ECGS?
Heparin comes as a powder in our lab. Sodium Salt. Just dissolve it in sterile dH2O and you're set..aliqot, freeze what you dont need.
Not sure what ECGS is so no comment on that.
Thanks Erik for the comment. that means I have to make a stock solution of 50 mg/ml and aliquote 1.12 mL into 500mL medium in order to get final concentration of 0.1 mg/ml heparin right?
ECGS is Endothelial Cell Growth Supplement.
You will need 25mg of ECGS at 0.05mg/mL for a 500mL media stock. You may prepare it using sterile PBS. Recommendations of the shelf-life of ECGS when added to the media differ, see what works best for you.
As far as the Heparin, I approached this differently. The recommendation is 180 USP units/mg. However, your bottle may have a different USP unit number, search for that number in the bottle. Here is some math to help with it:
(180 USP units/mg) x (0.1mg/ml) x (500ml) = 9,000 USP units needed for 500mL
(9,000 USP units) ÷ (X USP units/mg) = Y mg of Heparin Powder
Weigh the amount, and filter it using a 0.2um filter into your media.
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