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Our lab has purchased Antibiotic-Antimycotic (100X). How should I use it? Should I directly add 5 mL of Antibiotic-Antimycotic (100X) to 500 mL cell culture medium or should I dilute the Antibiotic-Antimycotic (100X) 100 times and then add 5 ml of the diluted Antibiotic-Antimycotic to 500 ml of cell culture medium? Product FAQ

Answer

The accurate way would be to dilute 5 mL of Antibiotic-Antimycotic (100X) into 495 mL of medium. Please note that the volume of medium in a 500 mL medium bottle is not exactly 500 mL. It could range from 500.5 - 510.5 mL.

Answer Id:: E17386

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Can I refrigerate or freeze the collected cell culture media for analysis later using the CyQUANT LDH Cytotoxicity Assay - Fluorescence Kit? Product FAQ

Answer

We do not recommend doing this. LDH is an enzyme that will lose activity in diluted form when stored over time, refrigeration would not limit this loss of activity and freezing would cause a greater loss of activity.

Answer Id:: E19029

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What is the best way to get a high yield of miRNA from exosomes isolated from cell culture media? Product FAQ

Answer

A pure sample of exosomes contains very low amounts of RNA. Therefore, the best way to increase yeild is to scale up the isolation. For cell culture media, you can try starting with 100 mL of sample.

Answer Id:: E7420

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I've purchased Gibco 2-Mercaptoethanol (Cat. No. 21985023). Should I add it to my cell culture medium right before every use or can I add it to the bulk medium in advance? Product FAQ

Answer

The 2-Mercaptoethanol must be added to the medium immediately before use because activity rapidly declines in diluted form. You can supplement enough medium needed for that day.

Answer Id:: E17388

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I would like to make a cDNA library for sequencing using mRNA isolated from exosomes. How much plasma or cell culture media would I need to use? Product FAQ

Answer

We would recommend using 20-50 mL of cell culture medium and 4-8 mL of serum or plasma. You can use the Total Exosome Isolation reagent to recover the exosomes. Ultracentrifugation is also an option, but yields will be lower.

Answer Id:: E7431

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Can my exosome sample (isolated from cell culture media or serum) be used in Western blotting directly? Product FAQ

Answer

Please see our protocols, including Western blotting:

- Schageman J., Zeringer E., Li M., Barta T., Lea K., Jian Gu, Magdaleno S., Setterquist R., and Vlassov A.V. The Complete Exosome Workflow Solution: From Isolation to Characterization of RNA Cargo. BioMed Research International. 2013, 2013:253957. doi: 10.1155/2013/253957.
- Zeringer E., Li M., Barta T., Schageman J., Pedersen K.W., Neurauter A., Magdaleno S., Setterquist R., Vlassov A.V. (2013) Methods for the extraction and RNA profiling of exosomes. World J Methodology. 3, 11-18.

Answer Id:: E7422

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What is your recommended method for thawing L-Glutamine before adding to cell culture media? Product FAQ

Answer

We recommend thawing at 2-8 degrees C and then warming at 37 degrees C until the material goes into solution. Mix it as it warms up to spend minimal time at elevated temperatures before you aliquot it.

Answer Id:: E17373

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Application Note: Flow rate and throughput of Nalgene 0.1 micron Rapid-Flow PES filter units with cell culture media Product Literature

Light-dependent generation of reactive oxygen species in cell culture media. Citations & References

  • Authors: Grzelak A, Rychlik B, Bartosz G
  • Journal: Free Radic Biol Med (2001) 30:1418-1425
  • PubMed ID: 11390187
Catalog #

Can I freeze complete cell culture media after supplementation with GlutaMAX supplement? Product FAQ

Answer

We don't recommend freezing complete medium as it may precipitate upon thawing. We recommend that complete media containing L-Glutamine and serum be stored at 2-8 degrees C, protected from light and used within 4 weeks.

Answer Id:: E17456

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Advanced Granulation Technology (AGT(TM)). An alternate format for serum-free, chemically-defined and protein-free cell culture media. Citations & References

  • Authors: Fike R, Dadey B, Hassett R, Radominski R, Jayme D, Cady D
  • Journal: Cytotechnology 2001 (36):33-39
  • PubMed ID: 19003312

StemPro® hESC SFM Human Embryonic Stem Cell Culture Medium Product Literature

Is there a fast way to reorder Gibco cell culture media? Product FAQ

Answer

Yes. If you are reordering the same formulation, in the same quantity and packaging, simply use the Quick Order link found at the top of every page of thermofisher.com, or visit your Order History page and click on the Reorder box.

Answer Id:: E11858

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Can proteins containing a His tag and secreted into tissue culture medium such as D-MEM (with serum and supplements) be directly purified on ProBond resin? Product FAQ

Answer

Proteins expressed in mammalian tissue culture media can be directly purified on ProBond resin, although not very effectively. D-MEM and other mammalian tissue culture media contain many contaminating proteins (and free amino acids like glutamine and histidine) that may compete for binding to the ProBond matrix. For best results, mammalian tissue culture media should be dialyzed (against binding buffer) or filtered prior to loading on equilibrated ProBond resin. Other methods such as ion-exchange or sizing columns can help to prepare the protein for effective isolation on the ProBond column. If necessary, the culture media may be adjusted to facilitate direct binding to the column. The pH of the media should be adjusted to 7.5 to 8.0, and the salt concentration in the media should be 0.5 M NaCl.

Answer Id:: E3663

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Application Note: Flow rate and throughput of cell culture media and serum using Nalgene Rapid-Flow filters with 0.2 micron PES membranes Product Literature

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