Reagent	Cat Number	FW	Final	For 1 LTR “1X”
1	Iscove’s MDM, powder	Gibco #12200-036	17.7 g/pk	1X = 1LTR	1 pkg
1*	NaHCO3	EM Sciences #SX0320-1	84.01 g/mol	36 mM	3.024 g
2	Hypoxanthine	Sigma #H9377	136.1 g/mol	1 mM	136 mg
3	Bathocuproine Disulfonate	Sigma #B125	564.5 g/mol	50 uM	28.2 mg
4	L-Cysteine • H2O	Fisher BP376-100	175.6 g/mol	1.5 mM	182 mg
5	2-mercaptoethanol	Sigma #M6250	14.3 M	0.2 mM	14.4 ul
6	Thymidine	Sigma #T9250	242.2 g/mol	0.16 mM	39 mg
7	Na-Pyruvate	Gibco #11360-070	100 mM	1 mM	10 ml
8	Pen/Strep (+ 29.2 mg/ml L-Gln) Stored at –20, thaw at 37 deg and invert several times to resuspend	Gibco #10378-016	10,000 u/ml	100 u/ml	10 ml

1. Dissolve pkg of IMDM in 800 ml warm MiliQ H₂O

2. Add 3.024 g NaHCO₃ while stirring at RT.

3. Add remaining ingredients while stirring at RT. Be sure all powders are dissolved.

4. If necessary, adjust pH to 7.4 with 1N NaOH.

5. Bring to 1L with H₂O

6. Filter Sterilize (0.22 um filter unit) and store at 2 – 8 deg C.

7. Add Heat-Inactivated Fetal Calf Serum to 15 % prior to use.