HEKPlus SFM (ATCC® ACS-4002)

Permits and Restrictions

View Permits

Product Format 1 L
Storage Conditions Store at 2°C to 8°C. Do not freeze. Protect from light.
Shipping Information Room Temperature OR 4C on Refrigerated Bricks
HEKPlus SFM should be supplemented with L-alanyl-glutamine or L-glutamine prior to use. The medium contains 0.1% Poloxamer 188, a surfactant shown to enhance cell yield and reduce cell aggregation. HEKPlus SFM also includes human transferrin and recombinant human insulin that have been shown to improve cell proliferation.

This medium has been optimized for use with ATCC HEK293T/17 SF suspension cells (ATCC ACS-4500) and GeneX Plus transfection reagent (ATCC ACS-4004) to reproducibly express a wide range of proteins at high levels. HEK293T/17 SF cells are pre-adapted to the medium and can be cultured in suspension and directly transfected in HEKPlus SFM using GeneXPlus transfection reagent. Transfection complexes can be prepared in the HEKPlus medium as well. In general, HEK293 cells currently adapted to serum-free medium can be subcultured directly using higher seeding densities. For cells currently adapted to serum containing medium, sequential adaptation to HEKPlus SFM is recommended.

Biosafety Level N/A
Introduction HEKPlus SFM is a xeno-free, serum-free, defined medium optimized for suspension culture and transfection of human embryonic kidney (HEK 293) cells. HEK293Plus SFM promotes long-term, high density cell growth and supports recombinant protein or adenovirus production in HEK293 cells.
Quality Control Specifications

Sterility tests are performed on each lot of medium using current USP methods. Each lot of medium is tested for the ability to support the growth of HEK 293 cell lines. Osmolality, pH, and endotoxin tests are also performed.







7.0 to 7.6




265 to 313 mOsm/kg H2O


Sterility Testing






≤ 1 EU/mL


Growth Promotion




*Please consult the Certificate of Analysis for lot-specific test results.

pH 7.0 to 7.6
Osmolality 265 to 313 mOsm/kg H2O
Endotoxin ≤ 1 EU/mL
Sterility Testing Pass
Notice: Necessary PermitsPermits

These permits may be required for shipping this product:

  • Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.
Basic Documentation
  1. HEKPlus Protein Expression System Parameters
    Cell density at transfection, ratio of DNA to GeneX
    Date Updated: 3/27/2014
  2. GeneXPlus transfection reagent
    Date Updated: 3/27/2014
  3. HEKPlus Protein Expression System vectors
    Expression vectors containing the SV40 origin of replication should be used. These vectors permit more plasmid copies to persist in the transiently transfected cells.
    Date Updated: 3/27/2014
  4. HEKPlus SFM supplements
    Complete culture medium is prepared by adding L-glutamine to the HEK
    Date Updated: 3/27/2014
  5. HEKPlus Protein Expression System time for optimal protein expression
    It usually takes 2 to 3 days to see optimal expression of intracellular proteins and 6 days for optimal expression of secreted proteins. It is recommended that a time course be performed for...
    Date Updated: 3/27/2014
  6. Advantage of using HEKPlus Protein Expression System
    The HEK
    Date Updated: 3/27/2014
  7. HEKPlus Protein Expression System advantages
    The HEK
    Date Updated: 3/27/2014
  8. HEKPlus Protein Expression System to express hard-to-express proteins
    Yes. The HEK
    Date Updated: 3/27/2014
  9. HEKPlus Protein Expression System Scale
    The HEK
    Date Updated: 3/27/2014
  10. HEKPlus Protein Expression System secreted proteins
    It is recommended to collect cell supernatants at day 6 post-transfection. However, one may want to perform a time course study to determine optimal time for collection post-transfection for each...
    Date Updated: 3/27/2014
  11. HEK293T/17 SF cells are clumping
    No. Small aggregates are easily dispersed with minimal pipetting and do not affect the performance of the cell line.
    Date Updated: 3/27/2014
  12. Replenishing medium for HEK293T/17 SF cells
    Yes. Incomplete media changes affect the cell viability and recovery. Spent medium should be completely removed and replaced with fresh medium every 2 to 3 days.
    Date Updated: 3/27/2014
  13. Media volume for HEK293T/17 SF cells
    For protein expression, it is important to have adequate shaking in the culture vessel. We usually add culture medium to 20% volume of a shaker flask.
    Date Updated: 3/27/2014