HIMEDIA Schneider’s Insect Medium w/ L-Glutamine
SKU: IM003-10L
HIMEDIA Schneider’s Insect Medium w/ L-Glutamine w/o Calcium Chloride and Sodium Bicarbonate 10L
Product Description:
Schneider's insect medium has been specially formulated for the in vitro culture of Drosophila melanogaster cells and tissues. A number of cell lines derived from Drosophila melanogaster are now available and are extensively used in genetic and molecular biology research. Drosophila cells are also used to study recombinant protein expression.
IM003 is Schneider's insect medium with L-glutamine. It does not contain calcium chloride and sodium bicarbonate, hence needs to be supplemented with calcium chloride and sodium bicarbonate while preparing the complete medium. The basal medium also needs to be supplemented with 5 - 20% of heat inactivated fetal bovine serum to supply the necessary growth factors to the insect cells. Schneider's insect medium is an extremely nutritious medium and supports rapid growth of cells derived from Drosophila. It can also be used to culture cell lines derived from other dipteran species. 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 |
|
|
Disodium hydrogen phosphate |
700.000 |
|
Magnesium sulphate heptahydrate |
3700.000 |
|
Potassium chloride |
1600.000 |
|
Potassium dihydrogen phosphate |
450.000 |
|
Sodium chloride |
2100.000 |
|
AMINO ACIDS |
|
|
Glycine |
250.000 |
|
L-Arginine hydrochloride |
483.810 |
|
L-Aspartic acid |
400.000 |
|
L-Cysteine hydrochloride |
78.000 |
|
L-Cystine dihydrochloride |
100.000 |
|
L-Glutamic acid |
800.000 |
|
L-Glutamine |
1800.000 |
|
L-Histidine hydrochloride monohydrate |
540.000 |
|
L-Isoleucine |
150.000 |
|
L-Leucine |
150.000 |
|
L-Lysine hydrochloride |
1650.000 |
|
L-Methionine |
800.000 |
|
L-Phenylalanine |
150.000 |
|
L-Proline |
1700.000 |
|
L-Serine |
250.000 |
|
L-Threonine |
350.000 |
|
L-Tryptophan |
100.000 |
|
L-Tyrosine disodium salt |
620.000 |
|
L-Valine |
300.000 |
|
ß-Alanine |
500.000 |
|
OTHERS |
|
|
Alpha-Ketoglutaric acid |
200.000 |
|
D(+) Glucose |
2000.000 |
|
D(+)-Trehalose |
2000.000 |
|
Fumaric acid |
100.000 |
|
L-Malic acid |
100.000 |
|
Succinic acid |
100.000 |
|
Yeast extract |
2000.000 |
Directions:
- Suspend 26.2gms in 900ml tissue culture grade water with constant, gentle stirring. Material will not go completely in the solution. Do not heat the water.
- Add 0.4gms of sodium bicarbonate powder (TC230) or 5.3ml of 7.5% sodium bicarbonate solution (TCL013) for each litre of the final volume of the medium being prepared. Stir until dissolved.
- Adjust the pH to 9.2 ± 0.2 with 1N sodium hydroxide with constant stirring. Solution may become turbid.
- Adjust the pH to 6.7 ± 0.2 with 1N HCl, with constant stirring. Solution will become clear.
- Add 0.6gms of anhydrous calcium chloride per litre of the medium. Prepare a solution by dissolving 0.6gms of anhydrous calcium chloride (TC097) in 50ml of tissue culture grade water. Add the solution slowly with constant stirring to avoid precipitate formation.
- Adjust the pH of the medium to 0.1-0.3 pH units using 1N NaOH or 1N HCl below the desired pH since it tends to rise during filtration. Make up the final volume to 1litre.
- Adjust the osmolality to 340 - 360mOsm/kg H2O. Osmolality can be increased by 10mOsm/kg H2O by adding 0.3gms per litre of 0.4gms per litre of potassium chloride (TC010) or sodium chloride (TC046). Osmolality can be decreased by 10mOsm/kg H2O by adding 27.8ml of water to per litre of the medium.
- Sterilize immediately by filtration using a membrane with porosity of 0.22 microns or less.
- 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