Technical Data
Media
Leibovitz L-15 Medium w/L-Glutamine, HEPES, Tris, Low Sodium (Powder)
L2100
Cell Culture Grade
Storage RT    Shipping RT
Components shown as mg/liter
Inorganic Salts:mg/liter
Calcium Chloride•2H2O185.45
Potassium Chloride400
Potassium Phosphate mono60
Magnesium Chloride93.65
Magnesium Sulfate97.67
Sodium Chloride6440
Sodium Phosphate dibasic190
Amino Acids:
L-Alanine225
L-Arginine500
L-Asparagine250
L-Cysteine120
L-Glutamine300
Glycine200
L-Histidine250
L-Isoleucine125
L-Leucine125
L-Lysine75
L-Methionine75
L-Phenylalanine125
L-Serine200
L-Threonine300
L-Tryptophan20
L-Tyrosine300
L-Valine100
Vitamins:
D-Pantothenic Acid•Ca1
Choline Chloride1
Folic Acid1
i-Inositol2
Nicotinamide1
Pyridoxine•HCI1
Riboflavin 5’ Phosphate(Na2)•H2O0.1
Thiamine Monophosphate•2H2O1
Other:
D-(+)-Galactose900
HEPES Free Acid (20mM)4766
Phenol Red10.2
Sodium Pyruvate550
TRIS (7mM)840
Total:17.8 g/L
The most commonly used buffering system for mammalian cell cultures is a bicarbonate/CO2 system, which requires CO2 regulators and incubators to supply a constant level of CO2. As a replacement, Leibovitz developed a bicarbonate-free medium, L15, with relatively high levels of certain amino acids in the free base form.

The most common method in use today for buffering mammalian cell cultures is the bicarbonate/CO2 system, which is based on the following equilibrium:
CO2 + H2O <-> HCO3" + H+

The use of bicarbonate is an attempt to mimic the buffering system of blood, but it has at least two major drawbacks. (1) The p/Q of bicarbonate is 6*1, which is far removed from the desired pH range of cell culture media (7-0-7-4). (2) Although bicarbonate is cheap, supplying a constant level of CO2 to cell cultures is definitely not, as it requires expensive CO2 incubators or fermentors.


Directions per Liter: Dissolve 17.8g in 800-900ml of ddH2O stirring gently until completely solubilized. Adjust pH of the medium to the desired level. Add additional water to bring the solution to1L If required, add sodium bicarbonate with stirring after solution has cooled. Filter-sterilize using a 0.22 micron membrane filter. Aliquot into sterile containers. Do not autoclave. Contains heat-labile compounds that can be damaged with autoclaving.

Storage and Stability: Store powdered media at RT. Opened bottles should be capped tightly and kept in a dark, low humidity environment. Prepared media should be kept at 4°C and used within a short period of time.


Important Note: This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.

1. Bernt, E. & Bergmeyer, H. U. (1974) Bergmeyer), p. 1304. Deerfield Beach, FL: Verlag Chemie Intl.
2. Eagle, H., Barban, S., Levy, M. & Schulze, H. O. (1958). The utilization of carbohydrates by
human cell cultures.. Biol. Chem. 233, SS1-558.
3. D. Barngrover, J. Thomas and W. G. Thilly, J. Cell Set. 78, 173-189 (1985) 173
4. Leibovitz, A. (1963) Amer. J. Hyg. 78:173-180.