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HIMEDIA RPMI- 1640 w/ L-Glutamine, Sodium Pyruvate, 4.5gms Glucose Per Litre and 10mM HEPES Buffer w/o Sodium Bicarbonate 10X1L, 50L

HIMEDIA RPMI- 1640 w/ L-Glutamine, Sodium Pyruvate

SKU: AT162-10X1L

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HIMEDIA RPMI- 1640 w/ L-Glutamine, Sodium Pyruvate, 4.5gms Glucose Per Litre and 10mM HEPES Buffer w/o Sodium Bicarbonate 10X1L, 50L

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.
AT162 is RPMI-1640 medium supplemented with Lglutamine, sodium pyruvate, 4.5gms glucose per liter and 10mM HEPES buffer. 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

1000.000

Magnesium sulphate anhydrous

48.840

Potassium chloride

400.000

Sodium chloride

6000.000

Sodium phosphate dibasic anhydrous

800.000

Sodium pyruvate

110.000

AMINO ACIDS

Glycine

10.000

L-Arginine hydrochloride

241.850

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

20.260

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

24.850

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

4500.000

Glutathione reduced

1.000

HEPES Buffer

2383.000

Phenol red sodium salt

5.300

 

Directions :

  1. Suspend 15.4gms in 900 ml tissue culture grade water with constant, gentle stirring until the powder is completely dissolved. Do not heat the water.
  2. 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.
  3. 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.
  4. Make up the final volume to 1000ml with tissue culture grade water.
  5. 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.
  6.  Aseptically add sterile supplements as required and dispense the desired amount of sterile medium into sterile containers.
  7. Store liquid medium at 2-8°C and in dark till use.