DMEM:Ham's F12 (1:1), w: 3.1 g/L Glucose, w: 1.6 mM L-Glutamine, w: 15 mM HEPES, w: 1.0 mM Sodium pyruvate, w: 1.2 g/L NaHCO3
DMEM:Ham's F12 is a widely recognized and extensively utilized basal medium in cell culture for biological research. It serves as a fundamental source of nutrients for the growth of various mammalian cell lines, particularly when supplemented with Fetal Bovine Serum (FBS).
This unique formulation combines Dulbecco's Modified Eagle Medium (DMEM) and Ham's F-12 (Ham's Nutrient Mixture F-12) in a precise 1:1 ratio. The addition of L-glutamine further enhances its composition.
DMEM, derived from Eagle's Minimal Essential Medium (EMEM), offers an increased concentration of amino acids and vitamins compared to its predecessor. In contrast, Ham's F-12 is based on Ham's F-10 medium, providing a complementary set of essential components.
To support optimal cell growth, it is common practice to supplement DMEM:Ham's F12 with FBS at a typical concentration of 5-10%. This addition is necessary as the medium lacks growth hormones, lipids, and proteins crucial for cellular development.
DMEM:Ham's F12 incorporates a pH buffer system and is often supplemented with phenol red, a pH indicator. Cultured cells in DMEM:Ham's F12, or any medium utilizing the bicarbonate buffer system, require a controlled CO2 environment of 5-10% to maintain appropriate pH levels. Phenol red enables monitoring of pH changes from 6.2 (yellow) to 8.2 (red).
Quality Control
- pH = 7.2 +/- 0.02 at 20-25°C.
- Each lot has been tested for sterility and absence of mycoplasma and bacteria.
Maintenance
- Keep refrigerated at +2°C to +8°C in the dark. Freezing as well as warming up to +37°C minimize the quality of the product.
- Do not heat the medium to more than 37°C or use uncontrollable sources of heat (e.g., microwave appliances).
- If only a part of the medium is to be used, remove this amount from the bottle and warm it up at room temperature.
- Shelf life for any medium except for the basic medium is 8 weeks from the date of manufacture.
Composition
Components | mg/L | |
Inorganic Salts | Calcium chloride x 2H2O | 154,45 |
Iron(III)-nitrate x 9H2O | 0,05 | |
Iron(II)-sulfate x 7H2O | 0,42 | |
Potassium chloride | 311,83 | |
Copper(II)-sulfate x 5H2O | 0.001 | |
Magnesium chloride anhydrous | 28,57 | |
Magnesium sulfate | 48,85 | |
Sodium chloride | 6,999.50 | |
Sodium dihydrogen phosphate anhydrous | 54,35 | |
di-Sodium hydrogen phosphate | 70,98 | |
Zinc sulfate x 7H2O | 0,43 | |
Other Components | D(+)-Glucose anhydrous | 3,151.00 |
HEPES | 3,574.50 | |
Hypoxanthine | 2,04 | |
Linoleic acid | 0,04 | |
DL-68-Lipoic acid | 0.103 | |
Sodium pyruvate | 110,00 | |
Phenol red | 8,10 | |
Putrescin x 2HCl | 0.081 | |
Thymidine | 0,36 | |
Amino Acids | L-Alanine | 4,45 |
L-Arginine x HCl | 147,35 | |
L-Asparagine x H2O | 7,50 | |
L-Aspartic acid | 6,65 | |
L-Cystine x 2HCl | 31,29 | |
L-Cysteine x HCl x H2O | 17,56 | |
L-Glutamine | 365,00 | |
L-Glutamic acid | 7,35 | |
Glycine | 18,75 | |
L-Histidine x HCl x H2O | 31,48 | |
L-Isoleucine | 54,37 | |
L-Leucine | 58,96 | |
L-Lysine x HCl | 91,37 | |
L-Methionine | 17,24 | |
L-Phenylalanine | 35,48 | |
L-Proline | 17,27 | |
L-Serine | 26,25 | |
L-Threonine | 53,55 | |
L-Tryptophan | 9,02 | |
L-Tyrosine x 2Na x 2H2O | 55,81 | |
L-Valine | 52,66 | |
Vitamins | D-(+)-Biotine | 0,004 |
D-Calcium pantothenate | 2,12 | |
Choline chloride | 8,98 | |
Folic acid | 2,66 | |
myo-Inositol | 12,51 | |
Nicotinamide | 2,02 | |
Pyridoxine x HCl | 2,03 | |
Riboflavin | 0,22 | |
Thiamine x HCl | 2,17 | |
Vitamine B12 | 0,68 | |
NaHCO3 | 1,200.00 |