Cat. №: СС231-02
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The Medium 199 was developed for research of chicken fibroblasts. It used for the culture of a wide range of attached cells. The media are developed on Hanks’ Salts for short term...
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Medium 199 with Hanks's Salts powder, with glutamine, without bicarbonate

Medium 199 with Hanks's Balanced Salts, with glutamine, without bicarbonate, is a dry mixture of inorganic salts, amino acids, vitamins, glucose and phenolic red for liquid medium preparation. Requires additional sodium bicarbonate.

The Medium 199 was developed specifically for nutritional research of chicken fibroblasts. Today it has been used for the culture of a wide range of attached cells. The media are developed on Hanks’ Balanced Saltsfor short term use under ambient atmospheric conditions – not for CO2 incubation.

This product is intended for research purposes.

Preparation instructions:

  1. 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.
  2. Rinse original package with a small amount of water to remove all traces of powder.

  3. Add a sodium bicarbonate to solution. Stir until dissolved.

  4. 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.

  5. When the required pH is reached add additional water to bring the solution to final volume.

  6. Sterilize immediately by filtration using a membrane with a porosity of 0.22 µm (or less). Aseptically dispense medium into sterile container.

  7. The finished product should be stored at temperature of +2 to +8 °C in a dry place protected from light.

  8. 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 and supplements

 mg/l

CaCl2 x 2H2O

185,45

Fe(NO3)3 x 9H2O

0,72

KCl

400

KH2PO4

60

MgSO4 x 7H2O

200

NaCl

8000

Na2HPO4

48

Adenine Sulfate 2H2O

11

Guanine

0,24

Hypoxanthine

0,3

Тhymine

0,3

Uracyl

0,3

Xanthine

0,3

Adenosine 5, 3 phosphate x 2Na x 3H2O

1,2

Adenosine 5, monophosphate x 2Na x 6H2O

0,29

Cholesterol

0,20

2-D-oxy-D-ribose

0,95

D-glucose

1000

Glutathione

0,05

Sodium acetat × 3Н2О

82,94

Тwin 80

20

Phenolic Red

20

Amino acids

 

L-Alanine

25

L-Arginine

57,87

L-Aspartic Acid

30

L-Cysteine HCl

0,09

L-Cystine

19,94

L-Tyrosine

39,93

L-Glutamic Acid

75

Glycine

50

L-Histidine

16,28

L-Hydroxyproline

10

L-Isoleucine

40

L-Leucine

60

L-Lysine HCl

70

L-Methionine

15

L-Phenylalanine

25

L-Proline

40

L-Serine

25

L-Threonine

30

L-Tryptophan

10

L-Valine

25

Vitamins

 

Ascorbic acid

0,05

D-Ca-Pantothenate

0,01

Choline Chloride

0,5

i-Inositol

0,05

Nicotinic acid

0,025

Nicotinamide

0,025

P-aminobenzoic acid

0,05

Pyridoxal HCl

0,025

Pyridoxine HCl

0,025

Riboflavin

0,01

Thiamine HCl

0,01

Vitamin A Acetate (Retinol Acetate)

0,14

Vit-E (a-tokoph.phos.) Na salt

0,01

Menadion sodim bisulfate

0,017

Calcium-ferol

0,1

D-biotin

0,01

Folic acid

0,01

  • Medium: 199
  • Sterility -
  • Glutamine: With glutamine
  • Salts: Hank's
  • HEPES: -
  • Sodium Pyruvate -
  • Sodium Bicarbonate -
  • Documents: For research purposes
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