LS-ProteinTrans Reagent
Large-Scale transient transfection for high protein production yield with reproducible results
| Cat.-no |
Description |
Amount |
Price € |
Shop |
| B01500 |
ProteinTrans Reagent Starter Kit
This starter Kit contents: 1.5 ml ProteinTrans Reagent
+ 5 ml CHO-Trans-Blast
Suitable for 0.5-1 Liter of cell culture |
1,5 ml |
209.00 |
add |
| B03000 |
ProteinTrans Reagent Kit
Kit contents: 3 ml ProteinTrans Reagent
+ 10 ml CHO-Trans-Blast
Suitable for 1-2 Liter of cell culture |
3 ml |
379.00 |
add |
| B15000 |
ProteinTrans Reagent Kit
Kit contents: 15 ml ProteinTrans Reagent
+ 50 ml CHO-Trans-Blast
Suitable for 5-10 Liter of cell culture |
15 ml |
1699.00 |
add |
| B30000 |
ProteinTrans Reagent Kit
Kit contents: 30 ml ProteinTrans Reagent reagent
+ 100 ml CHO-Trans-Blast
Suitable for 10-20 Liter of cell culture |
30 ml |
3099.00 |
add |
| Cat.-no |
Description |
Amount |
Price € |
Shop |
| 10003 |
ProteinTrans Reagent
Suitable for 1-2 Liter of cell culture |
3 ml |
359.00 |
add |
| 10015 |
ProteinTrans Reagent
Suitable for 5-10 Liter of cell culture |
15 ml |
1599.00 |
add |
| 20030 |
ProteinTrans Reagent
Suitable for 10-20 Liter of cell culture |
30 ml |
2899.00 |
add |
| 00005 |
CHO-Trans-Blast |
5 ml |
135.00 |
add |
ProteinTrans Reagent with CHO-Blast: manual
Ensure High Yield Protein Expression
Transient transfection of suspension-growing cells represents an attractive alternative to costly, time-consuming and labour intensive stable transfection.
The ProteinTrans transfection Kit has been designed for large scale up transient transfection and allows high protein production yield with reproducible results.
The ProteinTrans transfection Kit is dedicated to achieve High Yield Protein Expression in mammalian cells. This Kit has been designed for maximum efficiency in HEK293 and CHO cells growing in suspension.
The ProteinTrans transfection Kit, is ideal for bioreactor, spinner or flasks.
• High yield recombinant protein production
• Suitable for both HEK293 and CHO cells growing in suspension
• Compatible with any synthetic or regular media used for protein production
• Completely animal origin free
Results
1.Data 1: Designed to achieve High Yield Protein Expression in suspension-growing cells (Fig.1)
2.Data 2: Outperforms competitor: Secreted and intracellular expressed protein (Fig.2a/2b)
3.Data 3: Easy and efficient scale-up (Fig.3)
Product listing
Designed to achieve High Yield Protein Expression in suspension-growing cells
Fig.1: ProteinTrans transfection kit achieves higher potein yield than popular commercial transfection reagents in CHO cells adapted in suspension.
Secreted Alkaline Phosphatase (SEAP) was produced by transient transfection using ProteinTrans Reagent transfection kit, LP or FHD transfection reagents according to manufacturer's instruction. Day3, 5 and 7 supernatants were collected and SEAP activity was measured.
Outperforms competitor: Secreted and intracellular expressed protein
Fig.2a: ProteinTrans outperforms commercial reagent specific for protein production in HEK293 cells adapted suspension.
Secreted Alkaline Phosphatase (SEAP) or β-Galactosidase (β-Gal) were produced by transient transfection using ProteinTrans or FSM reagent according to manufacturer's instructions. HEK293 cells were cultured in 30 ml in 125 ml shaken flasks.
Fig.2b: ProteinTrans Kit outperforms commercial reagent specific for protein production in CHO cells adapted suspension.
Secreted Alkaline Phosphatase (SEAP) were produced by transient transfection using ProteinTrans in addition with ProteinTrans-Blast or FSM reagent according to manufacturer's instructions. CHO cells were cultured in 30 ml in 125 ml shaken flasks.
Easy and efficient scale-up
Fig.3: Protein production in both CHO and HEK293 suspension cells can be easily scaled up by using ProteinTrans transfection kit.
Secreted Alkaline Phosphatase (SEAP) was produced by transient transfection using ProteinTrans transfection Kit.CHO and HEK293 cells were cultured in 30 ml or 300 ml of medium in 125 ml or 1000 mL shaken flasks respectively.
The ProteinTrans transfection kit includes ProteinTrans REAGENT and CHOTrans-Blast.
The ProteinTrans is a DNA transfection reagent.
The CHO-Trans-Blast improves gene expression.
All components are completely animal origin free.
Troubleshooting
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Problem
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Comments and suggestions
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Low protein expression
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1- Suboptimal transfection conditions. Optimize the transfection conditions as describe in chapter 4 using a positive control plasmid.
2- Cell density. A non-optimal cell density at the time of transfection can lead to low protein expression. Cells must be actively dividing at the time of transfection. About 24h before transfection pass the cells at 0.5-0.6 x 106 cells/mL for having a cell density of ~ 1 x 106 cells/mL at the transfection time. However optimum cell density may depend on the cell model used and must be optimized as describe in the chapter 4.
3- Cell culture conditions. 1) Cells cultured for too many passages (>20-25 passages) may become resistant to transfection. Use freshly thawed cells that have been passaged at least twice. 2) Improperly cultured cells could lead to poor protein yield. Ensure complete adaptation to suspension growth conditions (medium, agitation…) to have cells growing as single and a viability >90%. 3) The presence of contaminants (mycoplasma, fungi) alters considerably the transfection efficiency. Thaw a new batch of cells or use appropriate antibiotic to eliminate contamination.
4- DNA quality. DNA must be highly pure, free of contaminants (proteins, phenol, ethanol etc.) and endotoxins levels must be very low since they interfere with transfection efficiencies.
5- Type of promoter. Ensure that DNA promoter can be highly expressed in the cells to be transfected. Another cells or viral-driven reporter gene expression can be used as a control.
6- Medium used for preparing DNA / transfection reagent complexes. It is critical that serum-free medium or buffer (HBS, PBS) are used during the preparation of the complexes. Avoid any direct contact of pure PROTEINTRANS reagent and pure nucleic acid solution with the plastic surface.
7- Incubation time. The optimal time range between transfection and assay is generally 2-5 days. As the protein expression profile will depend on the plasmid used we recommend you to first perform a kinetic from day 1 to day 7 to evaluate the optimum incubation time.
8- Transfection reagent handling. Reagents should be properly stored and must have an ambient temperature and be vortexed prior to use.
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Cells growing improperly
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1- Thawing cells are unhealthy. 1) Keep cell stock in liquid nitrogen at a density of 1 x 107 viable cells/mL until thawing; 2) Use low passage cells to make your own stock, 3) Use a freezing medium containing 10% DMSO and 90% of compatible medium.
2- Improper culture conditions. 1) Suspension cell adaptation is a key step for optimal growing cell conditions. Ensure that the culture medium is suitable for suspension growth. 2) Monitor cell density to prevent cell clumping. We suggest a 2-3 days frequency for cells passage at a density of 0.1-0.2 x 106 viable cells/mL. Avoid density > 1.5 x 106 viable cells/mL. 3) Monitor shaking condition to avoid toxicity, cell clumping or medium foam formation. We suggest 125 rpm with an orbital shaker as a starting point. 4) A cell culture volume of 1/3 of the total volume of the flask used is recommended.
3- Concentration of PROTEINTRANS/nucleic acid too high. Overloading the system won’t systematically lead to higher protein production because too high DNA quantity (and so high PROTEINTRANS reagent quantity) could lead to cell toxicity. Optimize the transfection conditions as describe in chapter 4 using a positive control plasmid.
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