HIMEDIA Grace’s Insect Medium w/ L-Glutamine w/o Sodium Bicarbonate
SKU: IM001-10L
HIMEDIA Grace’s Insect Medium w/ L-Glutamine w/o Sodium Bicarbonate 10L
Product Description:
Grace's medium is a modification of the original formula developed by Wyatt. Grace (1962) improved the medium by introducing ten water soluble vitamins in Wyatt's formula. Grace successfully established four strains of cells from ovarian tissue of Australian emperor gum
moth using this medium. These were the first continuous cell lines developed from insect tissue. IM001 is Grace's Insect Medium with L-glutamine. It can be used to culture cells derived from a variety of insects especially Lepidopterans and some species of Dipterans. Originally the basal medium was supplemented with plasma from insects, the tissues of
which were cultured or plasma from silkworm, Bombyx mori. Presently, the medium is supplemented with 5- 20% fetal bovine serum. 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 chloride dihydrate |
1320.000 |
|
Magnesium chloride anhydrous |
1068.200 |
|
Magnesium sulphate anhydrous |
1356.650 |
|
Potassium chloride |
1356.650 |
|
Sodium phosphate monobasic |
1007.000 |
|
AMINO ACIDS |
|
|
DL-Serine |
1100.000 |
|
DL-Serine |
650.000 |
|
L-Alanine |
225.000 |
|
L-Arginine hydrochloride |
700.000 |
|
L-Asparagine monohydrate |
400.000 |
|
L-Aspartic acid |
350.000 |
|
L-Cystine dihydrochloride |
25.000 |
|
L-Glutamic acid |
600.000 |
|
L-Glutamine |
600.000 |
|
L-Histidine hydrochloride monohydrate |
3376.000 |
|
L-Isoleucine |
50.000 |
|
L-Leucine |
75.000 |
|
L-Lysine hydrochloride |
625.000 |
|
L-Methionine |
50.000 |
|
L-Phenylalanine |
150.000 |
|
L-Proline |
350.000 |
|
L-Threonine |
175.000 |
|
L-Tryptophan |
100.000 |
|
L-Tyrosine disodium salt |
72.000 |
|
L-Valine |
100.000 |
|
ß-Alanine |
200.000 |
|
VITAMINS |
|
|
Choline chloride |
0.200 |
|
D-Biotin |
0.010 |
|
D-Ca- Pantothenate |
0.020 |
|
Folic acid |
0.020 |
|
Niacin |
0.020 |
|
Pyridoxine hydrochloride |
0.020 |
|
Riboflavin |
0.020 |
|
Thiamine hydrochloride |
0.020 |
|
myo-Inositol |
0.020 |
|
p-Amino benzoic acid (PABA) |
0.020 |
|
OTHERS |
|
|
Alpha-Ketoglutaric acid |
370.000 |
|
D(+) Glucose |
700.000 |
|
D-Fructose |
400.000 |
|
Fumaric acid, free acid |
55.000 |
|
L-Malic acid, free acid |
670.000 |
|
Succinic acid |
60.000 |
|
Sucrose |
26680.000 |
Directions:
- Suspend 45.9gms in 900ml tissue culture grade water with constant, gentle stirring until the medium is completely dissolved. Do not heat the water.
- Add 0.35gms of sodium bicarbonate powder (TC230) or 4.7ml of 7.5% of sodium bicarbonate solution (TCL013) for each litre of the medium. Stir until dissolved.
- Adjust the pH to 6.2 using 1N KOH.
- Make up the final volume to 1000ml
- Adjust the Osmolality as desired. For Lepidopterans cell line, osmolality of 340 - 360mOsm/KgH2O is recommended. The osmolality can be increased by 10mOsm/KgH2O by adding 0.4gms of potassium chloride (TC010) or 0.3gms of sodium chloride (TC046) to each litre of the medium. Osmolality can be decreased by 10mOsm/KgH2O by adding 27.8ml of water to per litre of medium.
- Sterilize the medium using a membrane filter 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.