RPMI-1640 medium powder, without glutamine, without bicarbonate
RPMI-1640 is a dry mixture of inorganic salts, amino acids, vitamins, glucose and phenol red, for liquid medium preparation. Requires additional sodium bicarbonate and glutamine.
The media are used for a wide range of mammalian cells including human myeloma, mouse hybridoma, human leukocytes and B and T lymphocytes. It was originally formulated for suspension and monolayer cultures of human leukemic cells.
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 and glutamine* 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 |
mg/l |
|
Ca(NO3)2 × 4H2O |
100 |
|
KCl |
400 |
|
MgSO4 x 7H2O |
100 |
|
NaCl |
6 000 |
|
Na2HPO4 |
800 |
|
D-glucose |
2000 |
|
Glutathione |
1 |
|
Amino acids |
|
|
L-Cystine |
49.86 |
|
L-Tyrosine |
19.96 |
|
L-Arginine |
200 |
|
L-Asparagine H2O |
56.82 |
|
L-Aspartic Acid |
20 |
|
L-Glutamic Acid |
20 |
|
Glycine |
10 |
|
L-Histidine |
15 |
|
L-Hydroxyproline |
20 |
|
L-Isoleucine |
50 |
|
L-Leucine |
50 |
|
L-Lysine HCl |
40 |
|
L-Methionine |
15 |
|
L-Phenylalanine |
15 |
|
L-Proline |
20 |
|
L-Serine |
30 |
|
L-Threonine |
20 |
|
L-Tryptophan |
5 |
|
L-Valine |
20 |
|
Vitamins |
|
|
D-Ca-Pantothenate |
0.25 |
|
Choline Chloride |
3 |
|
i-Inositol |
35 |
|
Nicotinamide |
1 |
|
Pyridoxine HCl |
1 |
|
Riboflavin |
0.2 |
|
Thiamine HCl |
1 |
|
P-aminobenzoic acid |
1 |
|
B12 |
0.005 |
|
D-biotin |
0.2 |
|
Folic acid |
1 |
|
Supplement media |
|
|
Phenolic red |
5 |
