HIMEDIA RPMI-1640 w/ L-Glutamine and 25mM HEPES
SKU: AT060-10X1L
HIMEDIA RPMI-1640 w/ L-Glutamine and 25mM HEPES buffer w/o Sodium Bicarbonate 10X1L
Product Description :
Roswell Park Memorial Institute (RPMI) media are a series of media developed by Moore et al for the culture of human normal and neoplastic cells in vitro.RPMI-1640 is the most commonly used medium in the series. A modification of McCoy's 5a medium, the medium was specifically designed to support the growth of human lymphoblastoid cells in suspension culture. Presently the medium is extensively used for a wide range of anchorage dependant cell lines. The medium needs to be supplemented with 5-20% fetal bovine serum. The medium is also known to support growth of cells in the absence of serum.
AT060 is modified RPMI-1640 medium supplemented with 25mM HEPES buffer and L-glutamine. HEPES, a zwitterionic buffer having a pKa of 7.3 at 37ºC prevents the initial rise in pH that tends to occur at the initiation of a culture and increases the buffering capacity of the medium. Users are advised to review the literature for recommendations regarding medium supplementation and physiological growth requirements specific for different cell
lines.
Composition :
|
Ingredients |
mg/L |
|
INORGANIC SALTS |
|
|
Calcium nitrate tetrahydrate |
100.000 |
|
Magnesium sulphate anhydrous |
48.840 |
|
Potassium chloride |
400.000 |
|
Sodium chloride |
6000.000 |
|
Sodium phosphate dibasic anhydrous |
800.000 |
|
AMINO ACIDS |
|
|
Glycine |
10.000 |
|
L-Arginine hydrochloride |
241.000 |
|
L-Asparagine monohydrate |
50.000 |
|
L-Aspartic acid |
20.000 |
|
L-Cystine dihydrochloride |
65.200 |
|
L-Glutamic acid |
20.000 |
|
L-Glutamine |
300.000 |
|
L-Histidine hydrochloride monohydrate |
20.960 |
|
L-Hydroxyproline |
20.000 |
|
L-Isoleucine |
50.000 |
|
L-Leucine |
50.000 |
|
L-Lysine hydrochloride |
40.000 |
|
L-Methionine |
15.000 |
|
L-Phenylalanine |
15.000 |
|
L-Proline |
20.000 |
|
L-Serine |
30.000 |
|
L-Threonine (Allo free) |
20.000 |
|
L-Tryptophan |
5.000 |
|
L-Tyrosine disodium salt dihydrate |
28.830 |
|
L-Valine |
20.000 |
|
VITAMINS |
|
|
Choline chloride |
3.000 |
|
D-Biotin |
0.200 |
|
D-Ca-Pantothenate |
0.250 |
|
Folic acid |
1.000 |
|
Niacinamide |
1.000 |
|
Pyridoxine hydrochloride |
1.000 |
|
Riboflavin |
0.200 |
|
Thiamine hydrochloride |
1.000 |
|
Vitamin B12 |
0.005 |
|
i-Inositol |
35.000 |
|
p-Amino benzoic acid (PABA) |
1.000 |
|
OTHERS |
|
|
D-Glucose |
2000.000 |
|
Glutathione reduced |
1.000 |
|
HEPES Buffer |
5958.000 |
|
Phenol red sodium salt |
5.300 |
Directions :
- Suspend 16.4gms in 900ml tissue culture grade water with constant, gentle stirring until the powder is completely dissolved. Do not heat the water.
- It is necessary to lower the pH to 4.0 with 1N HCl to completely dissolve this product. After it has dissolved completely, the pH can be raised to 7.2 with 1N NaOH prior to the addition of sodium bicarbonate.
- Add 2.0gms of Sodium bicarbonate powder (TC230) or 26.67ml of 7.5% Sodium bicarbonate solution (TCL013) for 1 litre of medium and stir until dissolved.
- Adjust the pH to 0.2-0.3 pH units below the desired pH using 1N HCl or 1N NaOH since the pH tends to rise during filtration.
- Make up the final volume to 1000ml with tissue culture grade water.
- Sterilize the medium immediately by filtering through a sterile membrane filter with a porosity of 0.22 micron or less, using positive pressure rather than vacuum to minimize the loss of carbon dioxide.
- Aseptically add sterile supplements as required and dispense the desired amount of sterile medium into sterile containers.
- Store liquid medium at 2-8°C and in dark till use.