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Minimal / Rich Growth Media

Minimal Media Reagents

The E. coli expression system using minimal media is the most popular means to over-express isotope-enriched, recombinant protein for NMR investigations. The primary reason for the popularity of the use of minimal media is that it remains to be the
most economic media available to grow isotope-labeled bacteria. Minimal media is a mixture M9 salts, glucose (as the sole carbon source) and ammonium chloride or ammonium sulfate (as the sole nitrogen source) that is formulated in either H2O or D2O. Often, depending on the E. coli strain used, additional reagents, such as cofactors, are added to the media to enhance growth.

CK Gas Isotopes is happy to be able to offer CIL’s minimal media reagents (*denotes products kept in stock within UK and available for next day delivery):

  • CLM-1396 – D-Glucose (U-13C6, 99%) *
  • DLM-2062 – D-Glucose (1,2,3,4,5,6,6-D7, 98%)
  • CDLM-3813 – D-Glucose (U-13C6, 99%; 1,2,3,4,5,6,6-D7, 97-98%)
  • CLM-1510 – Glycerol (13C3, 99%)
  • DLM-558 – Glycerol (D8, 99%)
  • NLM-467 – Ammonium chloride (15N, 99%) *
  • NLM-713 – Ammonium sulfate (15N2, 99%) *
  • DLM-4-99 – Deuterium oxide (D, 99%)
  • DLM-4-99.8 – Deuterium oxide (D, 99.8%)
  • DLM-4-1L – Deuterium oxide (D, 9 9%) *

Please click here for additional info

Rich E.coli Media 

Although growth in minimal (M9) media is economical, there is no substitute for the enhanced growth rates and increased levels of protein expression that may be gained by the use of a rich medium. Rich bacterial media are complex formulations that are usually derived from algal hydroylsates and contain all the necessary nutrients to promote excellent growth. CIL offers a number of rich media used in labeled protein expression using bacterial systems.

BioExpress® 1000
BioExpress® 1000 is CIL’s all-time classic rich bacterial cell growth medium. BioExpress® 1000 provides excellent growth and expression characteristics for a number of different bacterial systems. BioExpress® 1000 contains nearly the same level of amino acids as LB medium. Glucose levels range from 0.1-0.5 g / L, depending on the batch. BioExpress® 1000 media is prepared by adding sterile cell-culture grade water and mixing. Please note that D2O is required for reconstitution for products containing deuterium. BioExpress® 1000 is supplied as a 100 mL sterile liquid 10x concentrate, and reconstitutes to 1 L with no final pH adjustment required; 10 mL sample sizes are also available. The 10 mL sample size reconstitutes to make 100 mL of media
with no final pH adjustment required.

 

  • CGM-1000-C BioExpress® 1000 (U-13C, 98%) (10x Concentrate)
  • CGM-1000-D BioExpress® 1000 (U-D, 98%) (10x Concentrate)
  • CGM-1000-N BioExpress® 1000 (U-15N, 98%) (10x Concentrate)
  • CGM-1000-CD BioExpress® 1000 (U-13C, 98%; U-D, 98%) (10x Concentrate)
  • CGM-1000-CN BioExpress® 1000 (U-13C, 98%; U-15N, 98%) (10x Concentrate)
  • CGM-1000-DN BioExpress® 1000 (U-D, 98%; U-15N, 98%) (10x Concentrate)
  • CGM-1000-CDN BioExpress® 1000 (U-13C, 98%; U-15N, 98%; U-D, 98%) (10x Concentrate)
  • CGM-1000-U BioExpress® 1000 Unlabeled (10x Concentrate)

 

Celtone® Complete
Celtone® Complete yields a growth rate comparable to LB media, allowing for inoculation and induction within one working day. Glucose levels range from ~0.3 to 0.6 g / L, depending on the batch. Celtone® Complete is a ready-to-use medium that does not require dilution or pH adjustment. Each lot is tested for sterility, cell growth and protein expression. Celtone® Complete is available in 0.1 L and 1 L sizes.

  • CGM-1040-C Celtone® Complete Medium (13C, 98%+)
  • CGM-1040-D Celtone® Complete Medium (D, 97%+)
  • CGM-1040-N Celtone® Complete Medium (15N, 98%+)
  • CGM-1040-CN Celtone® Complete Medium (13C, 98%+; 15N, 98%+)
  • CGM-1040-DN Celtone® Complete Medium (D, 97%+; 15N, 98%+)
  • CGM-1040-CDN Celtone® Complete Medium (13C, 98%+; D, 97%+; 15N, 98%+)
  • CGM-1040-U Celtone® Complete Medium (Unlabeled)

 

Celtone® Powder
Celtone® Powder is CIL’s most flexible nutrient-rich media. The advantage of Celtone® Powder is that researchers can formulate a custom medium based on their specific research needs. Depending on cell line and desired performance, this powdered media can be used at concentrations ranging from 1 g to 10 g per liter. Truly exceptional performance has been achieved using 10 g of Celtone® Powder in 1 L of medium containing M9 salts, 2-3 g / L glucose and 1 g of ammonium chloride. Because it is a powder, this product has the longest shelf life of any fully rich bacterial cell growth medium. Please note that if deuterium labeling is desired, D2O must be used in media
preparation. Also note that it is normal to have insoluble material present after dissolution. This material may be removed using filter paper prior to sterile filtration and will not affect performance of the medium. Celtone® Powder is available in 0.5 g and 1 g packaged sizes.

  • CGM-1030P-C Celtone® Base Powder (13C, 98%+)
  • CGM-1030P-D Celtone® Base Powder (D, 97%+)
  • CGM-1030P-N Celtone® Base Powder (15N, 98%+)
  • CGM-1030P-CN Celtone® Base Powder (13C, 98%+; 15N, 98%+)
  • CGM-1030P-DN Celtone® Base Powder (D, 97%+; 15N, 98%+)
  • CGM-1030P-CDN Celtone® Base Powder (13C, 98%+; D, 97%+; 15N, 98%+)
  • CGM-1030P-U Celtone® Base Powder (Unlabeled)

Silantes® E. coli-OD2
Silantes® E. coli-OD2 is made from bacterial hydroylsate and primarily contains amino acids, some low MW oligopeptides and almost no carbohydrates (~0.1 g of glucose / L). The bacterial strain used is a chemolithoautotrophic organism which grows on inorganic substrates that are isotopically labeled. Silantes® OD2 media are ready-to-use sterile solutions and can be used instantly for the fermentation of bacteria. Silantes® E. coli-OD2 media is available in 0.2 L and 1 L sizes.

  • CGM-1020-SL-C E. coli-OD2 (Rich Growth Media for E. coli ) (13C, 98%)
  • CGM-1020-SL-D E. coli-OD2 (Rich Growth Media for E. coli ) (D, 98%)
  • CGM-1020-SL-N E. coli-OD2 (Rich Growth Media for E. coli ) (15N, 98%)
  • CGM-1020-SL-CN E. coli-OD2 (Rich Growth Media for E. coli ) (13C, 98%; 15N, 98%)
  • CGM-1020-SL-DN E. coli-OD2 (Rich Growth Media for E. coli ) (D, 98%; 15N 98%)
  • CGM-1020-SL-CDN E. coli-OD2 (Rich Growth Media for E. coli ) (13C, 98%; D, 98%; 15N, 98%)
  • CGM-1020-SL-U E. coli-OD2 (Rich Growth Media for E. coli ) (Unlabeled)

Spectra 9 Media
Spectra 9 Media is not a fully rich medium, however, it represents a cost-effective medium for E. coli growth and protein expression. It is comprised of labeled salts, labeled carbohydrates (>2 g glucose / L), and is supplemented with Celtone® Powder at a concentration of 1 g / L.

  • CGM-3030-C Spectra 9 (13C, 98%)
  • CGM-3030-D Spectra 9 (D, 97%+)
  • CGM-3030-N Spectra 9 (15N, 98%)
  • CGM-3030-CN Spectra 9 (13C, 98%; 15N, 98%)
  • CGM-3030-DN Spectra 9 (D, 97%+; 15N, 98%+)
  • CGM-3030-CDN Spectra 9 (13C, 98%; D, 97%+; 15N, 98%)
  • CGM-3030-U Spectra 9 (Unlabeled)

Yeast Media and Reagents

The over-expression of protein in yeast cells represents a powerful expression system for the source of properly folded and functional eukaryotic protein. Please see CIL Application Note 26 (page 71) for additional information regarding yeast as a viable expression system to produce isotope-enriched protein for NMR investigations.

Silantes® rich Yeast-OD2 Media for yeast are ready-to-use formulations. The problem of optimizing standard minimal media containing glucose or methanol as a sole carbon source therefore does not arise when using Yeast-OD2 growth media. For yeast that grows well in minimal media (e.g. pichia), CIL offers three different single- and double-labeled carbon sources in 15N ammonium salts. Silantes® rich Yeast-OD2 Media is available in a 1 L packaged size. Please see CIL Application Note 26 (page 71) for additional information regarding the use of methanol, glucose and ammonium salts for use in yeast

 

  • CGM-4020-SL-C Yeast-OD2 Growth Media (13C, 98%)
  • CGM-4020-SL-N Yeast-OD2 Growth Media (15N, 98%)
  • CGM-4020-SL-CN Yeast-OD2 Growth Media (13C, 98%; 15N, 98%)
  • CGM-4020-SL-U-S Yeast-OD2 Growth Media (Unlabeled)
  • NLM-467 Ammonium chloride (15N, 99%)
  • NLM-713 Ammonium sulfate (15N2, 99%)
  • CLM-1396 D-Glucose (13C6, 99%)
  • DLM-2062 D-Glucose (U-D7, 98)
  • CDLM-3813 D-Glucose (U-13C6, 99%; U-D7, 98%)
  • CLM-1510 Glycerol (13C3, 99%)
  • DLM-558 Glycerol (U-D8, 99%)
  • CDLM-7745 Glycerol (U-13C3, 99%; U-D8, 98%)
  • CLM-359 Methanol (13C, 99%)
  • CDLM-1035 Methanol (13C, 99%; D3,98%)

Insect Cell Media

CK Gas Isotopes is able to offer CIL’s BioExpress® 2000, a rich growth media for culturing insect cells. The use of BioExpress® 2000 in the uniform 15N and selective amino acid-type labeling of the catalytic domain of AbI kinase is described in CIL Application Note 14. The use of Experimental Design in the optimization of protein yield using BioExpress® 2000 is exemplified in CIL Application Note 20. BioExpress® 2000 is packaged as two components: a solid powder (a proprietary blend of inorganic salts, carbohydrates, and labeled amino acids) and a liquid component (fatty acid solution). Selective amino acid-type labeling is possible with BioExpress® 2000 because the amino acid content is chemically defined.

  • CGM-2000-N BioExpress® 2000 (U-15N, 98%)
  • CGM-2000-N-S BioExpress® 2000 (U-15N, 98%)
  • CGM-2000-CN BioExpress® 2000 (U-13C, 98%; U-15N, 98%)
  • CGM-2000-U BioExpress® 2000 (Unlabeled)
  • CGM-2000-U-S BioExpress® 2000 (Unlabeled)
  • CGM-2000-CUSTOM BioExpress® 2000 (Labeled amino acids to be specified by customer at time of request)

Mammalian Cell Media

Mammalian Cell Media
There is continued interest in obtaining labeled recombinant protein from mammalian cells because eukaryotic protein expressed in mammalian cells has the greatest probability of being properly folded and functional. CIL offers the only commercially available labeled mammalian media intended for the production of labeled protein with yields suitable for NMR studies. Similar growth characteristics should be obtained using BioExpress® 6000 as with using Dulbecco’s Modified Eagle Medium (DMEM). The amino acid content in BioExpress® 6000 is chemically defined so many different custom labeling strategies may be realized. For example, Harold Schwalbe, Karla Werner, and Judith Klein-Seetharaman at Goethe University have used BioExpress® 6000 to express rhodopsin from HEK293 cells that is labeled at the Gly, Lys, Leu, Gln, Ser, Thr, Val and Trp residues with either 15N or 13C,15N.

  • CGM-6000-N BioExpress® 6000 (U-15N, 98%)
  • CGM-6000-N-S BioExpress® 6000 (U-15N, 98%)
  • CGM-6000-CN BioExpress® 6000 (U-13C, 98%; U-15N, 98%)
  • CGM-6000-U-S BioExpress® 6000 (Unlabeled)
  • CGM-6000-CUSTOM BioExpress® 6000 (Labeled amino acids to be specified by customer at time of request)


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Wellington House,
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Ibstock,
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LE67 6HP

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Tel: 01530 267200
E-mail: enquiries@ckisotopes.com

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Tel: 01530 267209
E-mail: sales@ckgas.com

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