RPMI-1640 medium powder, without glutamine, without bicarbonate
RPMI-1640 is a dry mixture of inorganic salts, amino acids, vitamins, glucose and phenolic 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. Mix 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 cultural 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 in solution during long-term storage, so it can be added sterile glutamine directly before using the media. The stable analog of glutamine is alanyl-glutamine (Сat.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*4Н2O |
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-Aspargine 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 |
|
В12 |
0,005 |
|
D-biotin |
0,2 |
|
Folic acid |
1 |
|
Supplement media |
|
|
Phenolic red |
5 |
