Component	Final concentration	Stock	Amount
MTX	25 nM	1 mM	12.5 μl
MTX	50 nM	1 mM	25 μl
MTX	75 nM	1 mM	37.5 μl
MTX	100 nM	1 mM	50 μl
MTX	200 nM	1 mM	100 μl
MTX	300 nM	1 mM	150 μl
MTX	400 nM	1 mM	200 μl
MTX	500 nM	1 mM	250 μl
MTX	1 μM	1 mM	500 μl

Add the indicated amount of MTX to 500 ml Alpha MEM containing 5% FBS