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Amino Acids & Vitamins

By Corning® cellgro®

Amino Acids & Vitamins

L-Glutamine is an essential amino acid required by virtually all mammalian and insect cells grown in culture. Once deaminated, it is used as an energy source, incorporated into protein, and used in nucleic acid metabolism.

Undergoing spontaneous degradation, the breakdown of the amino acid results in the formation of pyrrolidone carbonic acid and potentially harmful ammonia. In addition, exposure to warm temperatures increases this intramolecular cyclization reaction.

A dipeptide derivative of L-glutamine, L-alanyl-L-glutamine incorporates L-alanine that protects the unstable alpha amino acid group. Aminopeptidases within the cells cleave the dipeptide, gradually releasing both L-glutamine and L-alanine for use by the cell.

Extremely stable, the dipeptide withstands autoclaving with minimal loss, while L-glutamine is almost completely destroyed when exposed to the same conditions.

Cat.No. Description Pack Size Shipped Storage Intended Use
23-25-015-CI L-Alanyl-L-Glutamine (Dipeptide L-glutamine), Liquid
200 mM Solution (with 8.5 g/L NaCl)
1 x 100 mL A R/A I
23-25-005-CI L-Glutamine, Liquid
200 mM Solution (29.23 mg/mL with 8.5 g/L NaCl)
6 x 100 mL F R I
23-25-005-CV L-Glutamine, Liquid
200 mM Solution (29.23 mg/mL with 8.5 g/L NaCl)
6 x 500 mL F R I
23-61-030-RM L-Glutamine, Powder
MW 146.15
1 x 100 g A R IVD
23-61-030-RO L-Glutamine, Powder
MW 146.15
1 x 500 g A R IVD
23-61-030-RR L-Glutamine, Powder
MW 146.15
1 x 1 kg A R IVD

Supplementation Table for L-Glutamine and L-Alanyl-L-Glutamine to L-Glutamine-Deficient Culture Media

AMEM 2 mM / 292 mg/L
BME 2 mM / 292 mg/L
DMEM 4 mM / 584 mg/L
DMEM/F-12 2.5 mM / 365.10 mg/L
EMEM 2 mM / 292 mg/L
Iscove's DMEM 4 mM / 584 mg/L
M199 0.68 mM / 100 mg/L
RPMI 1640 2 mM / 292 mg/L
Cat.No. Description Pack Size Shipped Storage Intended Use
23-25-030-CI MEM Amino Acids
50X Solution
without L-glutamine
6 x 100 mL A R I
23-25-025-CI MEM Nonessential Amino Acids
100X Solution
6 x 100 mL A R I
23-25-020-CI MEM Vitamins
100X Solution
6 x 100 mL F F I

F = -5 to -20°C
R = 2 to 8°C
A = 15 to 30°C
* = Glass
** = Flexible Media Bag
I = Sterile, IVD Product
NI = Sterile, Research Use Only, Not for IVD Use
IVD = In Vitro Diagnostic
Res = Research Use Only, Not for IVD Use

Catalog No. 23-25-030
50X Liquid
MEM Amino Acid Solution
Components mg/L
L-Arginine•HCl 6320.00
L-Cystine•2HCl 1560.00
L-Histidine•HCl•H2O 2095.00
L-Isoleucine 2625.00
L-Leucine 2625.00
L-Lysine•HCl 3625.00
L-Methionine 750.00
L-Phenylalanine 1625.00
L-Threonine 2380.00
L-Tryptophan 500.00
L-Tyrosine•HCl 2160.00
L-Valine 2340.00
Specifications
pH (before buffer) 1.50
pH (after buffer) 1.5±0.5
Osmolality (mOsm) INFO
Catalog No. 23-25-025
100X Liquid
MEM Nonessential
Amino Acid Solution
Components mg/L
L-Alanine 890.00
L-Asparagine•H2O 1500.00
L-Aspartic acid 1330.00
L-Glutamic acid 1470.00
Glycine 750.00
L-Proline 1150.00
L-Serine 1050.00
Specifications
pH (before buffer) 3.5±0.2
pH (after buffer) 3.5±0.5
Osmolality (mOsm) 74±15
Catalog No. 23-25-020
100X Liquid
MEM Vitamin Solution
Components mg/L
NaCl 8500.00
D-Calcium pantothenate 100.00
Choline chloride 100.00
Folic acid 100.00
i-Inositol 200.00
Nicotinamide 100.00
Pyridoxine•HCl 100.00
Riboflavin 10.00
Thiamine•HCl 100.00
Specifications
pH (before buffer) N/A
pH (after buffer) 6.8-7.6
Osmolality (mOsm) 250±50

L-Glutamine

L-Glutamine is an essential amino acid required by virtually all mammalian and insect cells grown in culture. It is more labile in cell culture solution than other amino acids. The rate and extent of L-glutamine degradation are related to storage temperature, age of the product, and pH. Mediatech guarantees media containing L-glutamine to be functional until the expiration date reported on the label and the Certificate of Analysis if properly stored.

For convenience, many customers prefer to use cell culture media containing L-glutamine, but Mediatech does offer media deficient in L-glutamine. L-Glutamine is offered as a 200 mM solution (29.23 mg/mL), Cat.No. 23-25-005, or as a powder (MW 146.15), Cat.No. 23-61-030, for supplementation.

For information regarding the suggested additions of L-glutamine to deficient media, please refer to the formulation tables.

Note: L-Glutamine degradation results in a build-up of ammonia which could have a deleterious effect on some culturing systems. Therefore, caution should be used when exceeding the originally formulated level of L-glutamine. For most cell types, ammonia toxicity will be a more important concern than the limitation of L-glutamine.

Avoid freeze-thaw cycles when using L-glutamine. If a stock solution is maintained, thaw the stock solution once, aliquot into smaller volumes, and freeze. This enables each aliquot to be thawed and used when needed without having to thaw the other aliquots, thereby avoiding freeze-thaw cycles.

Procedure: Preparation of L-Glutamine from Powder

  1. Examine the powder for caking, which may indicate moisture uptake and compromise the stability of L-glutamine.
  2. Prepare a stock solution from which working concentrations may easily be obtained. Determine the amount of L-glutamine powder and water necessary to make the stock solution; a 200 mM (29.23 g/L) solution is often a convenient choice due to its simple dilution to working concentrations.
  3. Use distilled, deionized water equilibrated to room temperature, 15-30°C; do not heat. Measure the appropriate amount of water for the desired volume of stock solution to be prepared; add the water to a clean, dry bottle for mixing.
  4. Add the powder slowly to the water and mix. This is most easily achieved by using a bottle with a screw cap. Ensure there are no clumps and the powder is completely in solution.
  5. You may wish to add a small amount of the solution to the weigh boat, or other weigh vessel, to rinse any remaining powder. Return this "wash" back to the mixing vessel. Ensure there are no clumps or visible powder in the solution.
  6. Sterilize by filtration through a sterile 0.22 micron filter into a sterile collection vessel.
  7. Dispense small aliquots into containers appropriate for freezing. This prevents the freeze-thaw process of large quantities when thawing for media supplementation or other use.
  8. Store the stock solution at -20°C (frozen). When needed, allow the stock solution to thaw in the refrigerator (2-8°C), usually overnight depending on the size of the aliquot.
  9. Determine the amount of the stock solution to supplement into the media to yield the desired working concentration.

Note: L-Glutamine may precipitate during the thawing process. The bottle may be placed in a 37°C water bath and gently swirled during thawing to get L-glutamine back into solution. Use caution when heating to avoid degradation of the amino acid. Do not leave the bottle in the water bath once the product has been thawed.

                   
Cat.No. Name Comments Pkg Size EA     PK    
23-25-005-CI L-glutamine, liquid 200mM solution 6-pack/100mL bottles       $106.06
1/PK
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23-25-005-CV L-glutamine, liquid 200mM solution 6-pack/500mL bottles       $394.43
1/PK
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23-25-020-CI MEM vitamins 100X solution 6-pack/100mL bottles       $73.10
1/PK
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23-25-025-CI MEM nonessential amino acids 100X solution 6-pack/100mL bottles       $79.68
1/PK
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23-25-030-CI MEM amino acids 50X solution w/o L-glutamine 6-pack/100mL bottles       $91.02
1/PK
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23-61-030-RM L-glutamine, powder MW 146.15 100-gram unit $89.23
1/EA
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23-61-030-RO L-glutamine, powder MW 146.15 500-gram unit $226.66
1/EA
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23-61-030-RR L-glutamine, powder MW 146.15 1kg unit $143.64
1/EA
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