Transient Transfection

In transient transfection, the introduced nucleic acid exists in the cell only for a limited period of time and is not integrated into the genome. Therefore, the new gene will not be replicated. The effects of the foreign gene within the cell last only a short amount of time (usually several days), after which the foreign gene is lost through cell division or other factors.

Cells that are frequently used for transient transfection are HEK293 or CHO cells. Large volume transient transfection of HEK293 and CHO cells adapted to suspension culture has addressed the need to obtain amounts of recombinant protein without the time-consuming and labor-intensive creation of stable cell lines.

Why Choose Transient Transfection?

  • Quickly transform cloned genes into large amounts of protein in 3-7 days.
  • Obtain fully post-translational modified and active mammalian protein.
  • Easily purify secreted protein from serum-free cell culture medium.
  • Expand culture volume from that of 50 multi-well plates to that of 25L and more.

Transient Transfection Expression

Whether you are focused on discovery, production of targets, or characterization of interesting gene products or antibodies, our one-stop transient transfection expression will help optimize the expression and purification of your target gene.

Service Process Deliverables Timeline
Gene synthesis
  • Codon optimization
  • Gene synthesis
  • Subcloning & Max-preparation
  • DNA sequencing report
  • Cloning vector
  • Expression results
  • QC data
2 weeks
Transient
Transfection
Expression
  • Pilot expression
  • Western blot analysis
2 weeks
  • Scale up expression (>200ml)
  • Purification (>90% purity)
  • QC: SDS-PAGE & Western blot
According to your requirements

Notes:

  1. You are only required to provide the gene sequence of protein/antibody and provide the cells used for transfection
  2. The volume of the culture medium to be amplified and expressed will be determined as per your request (>200ml)
  3. The purity of the delivered protein will be determined as per your request (>90%)
  4. Label selection: proteins are His-tagged by default. If other taggings are required, please contact us.
  5. QC approach is optional: SDS-PAGE, WB, MS, etc.
  6. Additional services: proteomics analysis, transportation requirements (buffer/freeze-drying)

General Guidelines for Transfection

  • Cell culture
  • First of all you need a cell line which is easy to transfect and will allow high product titers. Large volume transient transfection of HEK293 and CHO cells adapted to suspension culture has addressed the need to obtain high amounts of recombinant protein. Some transfection protocols require serum-free conditions for optimal performance, since serum can interfere with many commercially available transfection reagents.

  • Plasmid transformation & plasmid max-preparation
  • DNA quality strongly influences the results of transfection experiments. The best results are achieved when plasmid DNA of the highest purity is used for transfection.

  • Confluency & Transfection
  • Cells should be transfected at 50–80% confluency. Too few cells will cause the culture to grow poorly. Too many cells results in contact inhibition.

    Ensure that cells are active and at least 90% of them survive before transfection. The cell density is controlled at 1.5-2×106 cells/mL.

  • Transfection Technology
  • Several methods for transfection of foreign (target) gene have been developed using liposomes, calcium phosphate, electroporation, non-liposomal lipids, DEAE-dextran, microinjection and retroviral transfection.

    The selection of transfection technology can strongly affect the transfection efficiency. In ideal conditions, transfection should be quick and easy to operate, providing high efficiency and repeatable results with minimal cytotoxicity.

Contact Us

For quote, please fill in the online form and email to Sales@Detaibio.com.

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