Amino Acids:	Cat. Sigma #:	F.W.: (g/mol)	4 Liters
1. B-Alanine	A-7752	89.09	8,000 mg
2. DL-Alanine	A-7502	89.09	4,360 mg
3. L-Arginine	A-3909	210.7	1,760 mg
4. L-Asparagine • 1 H2O	A-8381	150.1	960 mg
5. L-Aspartic Acid	A-9256	133.1	440 mg
6. L-Cysteine-HCl H2O	Fisher BP-376	175.63	320 mg
7. L-Cystine	C-8767	240.3	120 mg
8. L-Glutamic Acid	G-1251	147.1	1,000 mg
9. L-Glutamine	G-1517	146.1	6,560 mg
10. Glycine	Gibco 1106-038	75.07	480 mg
11. L-Histidine	H-8000	155.2	640 mg
12. DL-Isoleucine	I-2627	131.2	360 mg
13. L-Leucine	L-8000	131.2	360 mg
14. L-Lysine	L-5626	182.7	600 mg
15. DL-Methionine	M-2768	149.2	800 mg
16. L-Phenylalanine	P-2126	165.2	800 mg
17. L-Proline	P-0380	115.1	27,600 mg
18. DL-Serine	S-4375	105.1	800 mg
19. L-Taurine	T-0625	125.1	1,080 mg
20. DL-Threonine	T-1520	119.1	400 mg
21. L-Tryptophan	Gibco 2110-010	204.0	400 mg
22. L-Tyrosine	T-3754	181.2	800 mg
23. DL-Valine	V-0375	117.2	840 mg

1. Mix 23 Amino Acids in ~2.6 L H₂O and stir at RT overnight to dissolve.

2. Mix independently, inorganic salts I, II and III in 280 ml, 160 ml and 160 ml H₂O, respectively.

3. Mix sugars plus organic acids in 560 ml.

4. Combine all ingredients, including vitamins.

5. Adjust pH to 7.4 with 10 N NaOH (approx. 5–8 ml/L) and bring to 4 L final volume.

6. Filter sterilize and store in 500 ml aliquots at 4 deg C.

7. Add HI-FCS to 10% before use.

^*Phenol red is optional and should be omitted if doing sedimentation assays.