Right now I am serum starving my cell lines. What is the amount of EGF I need to apply in per well of 6 well plate. What is the calculation for finding right amount of EGF?
i have already diluted my EGF with PBS.
Hello Md Abu Siddique
After starving your cell lines, you may use fresh serum-free medium as control, and you could use EGF concentration in the range of 0.5 ng/ml to 10ng/ml. You may prepare these working solutions from the EGF stock in either sterile PBS or serum-free media.
EGF comes in a lyophilized form. You may reconstitute with sterile distilled water to 1mg/ml (stock solution).
The stock solution can be further diluted with sterile PBS.
For instance, if you need 1ng/ml, you need to dilute the stock 1:1000 to get a diluted stock of 1ug/ml. Add 1ul of 1ug/ml (diluted stock) to 999ul of cell growth media (serum free media) to get 1ml of serum free media containing 1ng/ml of EGF. Mix well and add to the well of a 6-well plate.
Best.
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