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M2852-02 Medium 199 Modified w/ Hanks’, L-Glutamine w/o Cholesterol, Choline, Folic acid, Thymine (Powder, TC 199)

Specifications
References
Grade
Cell Culture Grade
EU Commodity Code
38210000
Shipping Temp
RT
Storage Temp
RT/4°C

In 1950, Morgan, Morton and Parker described one of the first totally defined media that did not depend largely on animal products or extracts as nutritive sources. This medium supported the growth of primary chick embryo heart cells. It has since become known as Medium 199. Its current use is with the addition of serum in virus and vaccine production, and in the culture of many non-transformed cells. It is also used in combination with less complex media. Medium 199 modified with Hanks’ Balanced Salts is designed for use with cells maintained in ambient (non-CO2) atmospheric conditions. M2852 differs from the original formulation by containing all L-amino acids. The original formulation contained 10 DL-amino acids, which are less biologically active on a per gram basis than the corresponding L-amino acids.

Appearance
Light tan to light orange, homogeneous, free flowing powder
Solubility
Orange, clear, complete
pH
As reported
Endotoxin
≤10EU/ml
Directions per Liter
Dissolve 10.772 grams per liter of distilled/deionized (DDI) water. Heat with stirring until completely solubilized. If required, add 2.20g sodium bicarbonate with stirring after solution has cooled. Adjust pH as necessary. Sterilize with 0.2um 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. Stable for 12 months after receipt. 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.
Media Formulation
Components shown as mg/liter
Inorganic Salts
Calcium Chloride•2H2O185.5
Ferric Nitrate•9H2O0.72
Potassium Chloride400
Potassium Phosphate mono60
Magnesium Sulfate•7H2O200.0
Sodium Chloride8000
Sodium Phosphate Dibasic47.5
AminoAcids
L-Alanine25
L-Arginine HCl70
L-Aspartic Acid30
L-Cysteine•HCI0.0987
L-Cystine•2Na•H2O25.16
L-Glutamic Acid66.82
L-Glutamine100
Glycine50
L-Histidine•HCl•H2O21.88
L-Hydroxyproline10
L-Isoleucine20
L-Leucine60
L-Lysine•HCI70
L-Methionine15
L-Phenylalanine25
L-Proline40
L-Serine25
L-Threonine30
L-Tryptophan10
L-Tyrosine•2Na•2H2O57.66
L-Valine25
Vitamins
L-Ascorbic Acid0.05
Biotin0.01
D-Pantothenic Acid•Ca0.01
Choline ChlorideAbsent
Calciferol0.1
FolicAcidAbsent
i-Inositol0.05
Menadione•NaHSO3•3H2O0.019
Nicotinamide (Niacinamide)0.025
Niacin (Nicotinic Acid)0.025
P-Aminobenzoic Acid0.05
Pyridoxal•HCI0.025
Pyridoxine•HCI0.025
Riboflavin0.01
Thiamine•HCI0.01
VitaminAAcetate0.1147
DL-Tocopherol PO4•Na2•2H2O0.0106
Other
Adenine Sulfate•2H2O10.98
ATP, Na2•3H2O1.098
5AMP•H2O (Adenylic Acid )0.2104
CholesterolAbsent
2-Deoxy-D-ribose0.5
Dextrose (D-Glucose)1000
Glutathione0.05
Guanine•HCl0.30
Hypoxanthine0.30
Phenol Red, Sodium17
D-Ribose0.5
Sodium Acetate50
ThymineAbsent
Tween 80®20
Uracil0.3
Xanthine0.3
References
1. Morgan, J.F., Morton, H.J., Parker, R.C., Proc. Soc. Exp. Biol. Med. 73: 1-8 (1950). 2. Morgan, J.F., Campbell, E., Morton, H.J., J.N.C.I. 16(2): 557-567 (1955). 3. Rutzky, L.P., Pumper, R.W., In Vitrol 9: 468 (1974). 4. Morton, H.J., Toinai, S., TCA Manual 4: 729 (1978). 5. Ryan, J.M., Walk, P.H., TCA Manual 5: 1043 (1979). 6. Bolton, W.E., Terrell, S.P., et al., Tissue Culture Methods 7: 39 (1982).
USBio References
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