Medium 199 with Hanks' Salts powder, with glutamine, without bicarbonate
Medium 199 with Hanks' Balanced Salts, with glutamine, without bicarbonate, is a dry mixture of inorganic salts, amino acids, vitamins, glucose and phenol red for liquid medium preparation. Requires additional sodium bicarbonate.
The Medium 199 was developed specifically for nutritional research of chicken fibroblasts. Today it has been used for the culture of a wide range of adherent cells. The media are developed on Hanks’ Balanced Salts for short term use under ambient atmospheric conditions – not for CO2 incubation.
This product is intended for research purposes.
Preparation instructions:
- Measure out 90% of final required volume of cell culture grade water. Water temperature should be 15-20°C. M ix gently, slowly add the powdered medium. Do not heat, stir until completely dissolved.
-
Rinse original package with a small amount of water to remove all traces of powder.
-
Add a sodium bicarbonate to solution. Stir until dissolved.
-
Measure pH after complete dissolution. The pH value should be lower than the finished solution pH near 0.2 units. To adjust pH, use cell culture-grade solutions of 1N HCl or 1N NaOH.
-
When the required pH is reached add additional water to bring the solution to final volume.
-
Sterilize immediately by filtration using a membrane with a porosity of 0.22 µm (or less). Aseptically dispense medium into sterile container.
-
The finished product should be stored at temperature of +2 to +8 °C in a dry place protected from light.
- Manipulate the product in aseptic conditions.
- Attention: after filtration pH increases by 0.1-0.3 units.
- Glutamine degrades during prolonged storage in solution. Sterile glutamine can therefore be added immediately before using the medium. Alanyl-glutamine is a stable glutamine analogue (Cat. No. Ф102626 or Ф005) available too.
Recommended Use:
- Do not use the product after its expiry date.
- Dry powder medium should be free flowing. Do not use if powder caked.
- Use gloves, mask and clothes adapted to the manipulation of the product to avoid contamination.
- Manipulate the product in aseptic conditions.
- Supplements such as antibiotics or L-Glutamine should be added as sterile supplements to the ready medium.
- Medium should be clear and free of opalescence and sediment.
HOW SUPPLIED: The powdered mix is available in ziplock bags or plastic cans.
SHELF LIFE: 3 years.
STORAGE: +2 to +8 °C. Protect the product from any form of humidity and light.
FORMULATION:
|
Salts and supplements |
mg/l |
|
CaCl2 x 2H2O |
185.45 |
|
Fe(NO3)3 x 9H2O |
0.72 |
|
KCl |
400 |
|
KH2PO4 |
60 |
|
MgSO4 x 7H2O |
200 |
|
NaCl |
8000 |
|
Na2HPO4 |
48 |
|
Adenine Sulfate 2H2O |
11 |
|
Guanine |
0.24 |
|
Hypoxanthine |
0.3 |
|
Thymine |
0.3 |
|
Uracyl |
0.3 |
|
Xanthine |
0.3 |
|
Adenosine 5, 3 phosphate x 2Na x 3H2O |
1.2 |
|
Adenosine 5, monophosphate x 2Na x 6H2O |
0.29 |
|
Cholesterol |
0.20 |
|
2-D-oxy-D-ribose |
0.95 |
|
D-glucose |
1000 |
|
Glutathione |
0.05 |
|
Sodium acetate × 3H2O |
82.94 |
|
Tween 80 |
20 |
|
Phenol Red |
20 |
|
Amino acids |
|
|
L-Alanine |
25 |
|
L-Arginine |
57.87 |
|
L-Aspartic Acid |
30 |
|
L-Cysteine HCl |
0.09 |
|
L-Cystine |
19.94 |
|
L-Tyrosine |
39.93 |
|
L-Glutamic Acid |
75 |
|
Glycine |
50 |
|
L-Histidine |
16.28 |
|
L-Hydroxyproline |
10 |
|
L-Isoleucine |
40 |
|
L-Leucine |
60 |
|
L-Lysine HCl |
70 |
|
L-Methionine |
15 |
|
L-Phenylalanine |
25 |
|
L-Proline |
40 |
|
L-Serine |
25 |
|
L-Threonine |
30 |
|
L-Tryptophan |
10 |
|
L-Valine |
25 |
|
Vitamins |
|
|
Ascorbic acid |
0.05 |
|
D-Ca-Pantothenate |
0.01 |
|
Choline Chloride |
0.5 |
|
i-Inositol |
0.05 |
|
Nicotinic acid |
0.025 |
|
Nicotinamide |
0.025 |
|
P-aminobenzoic acid |
0.05 |
|
Pyridoxal HCl |
0.025 |
|
Pyridoxine HCl |
0.025 |
|
Riboflavin |
0.01 |
|
Thiamine HCl |
0.01 |
|
Vitamin A Acetate (Retinol Acetate) |
0.14 |
|
Vit-E (a-tokoph.phos.) Na salt |
0.01 |
|
Menadion sodim bisulfate |
0.017 |
|
Calcium-ferol |
0.1 |
|
D-biotin |
0.01 |
|
Folic acid |
0.01 |
