ScreenFect(TM)UP-293

*Use ScreenFect™ UP-293 Dilution Buffer provided with this kit.

ScreenFect™ UP-293 is a transfection reagent kit which is highly efficient for production of antibodies and proteins in Expi293F™ or FreeStyle™ 293-F cells. ScreenFect™ UP-293 Booster, which is added to transfected cells, enhances remarkably protein expression level in cells.

• Low-priced
• High yield protein expression
• Just change your transfection reagent

Comparison of luciferase expression level in Expi293F™ Cells

This figure compares transfection reagent performance in Expi293F™ cells. Cells were transfected with a secreted Luc expression vector using commercially available transfection reagents and cultured in 30 mL of Expi293™ Expression Medium. 96 hours after transfection, cell supernatants were measured for emission intensity by Luciferase assay.

ScreenFect™ UP-293 achieves the highest luciferase expression level in comparison to other commercially available transfection reagents.

Expi293™, Freestyle™, Expi293F™ are registered trademarks of Thermo Fisher Scientific.

High production yield for IgG in Expi293F™ Cells

Heavy and light chains expression vectors were transfected using commercially available transfection reagent in Expi293F™ Cells. The Expi293F™ Cells were cultured in 30 mL of Expi293™ Expression Medium. 120 hours after transfection, supernatants were collected and electrophoresed. IgG was measured by CBB staining.

ScreenFect™ UP-293 has the same capabilities at other company's product.

Comparison of luciferase expression level in FreeStyle™ 293-F Cells

This figure compares transfection reagent performance in FreeStyle™ 293-F Cells. Cells were transfected with a secreted Luc expression vector using commercially available transfection reagent and cultured in 30 mL of FreeStyle™ 293 Expression Medium. Emission intensity of cell supernatants was measured by Luciferase assay, 96 hours post-transfection.

ScreenFect™ UP-293 provides considerably higher expression level than Product C.

Expi293™, Freestyle™, Expi293F™ are registered trademarks of Thermo Fisher Scientific.

Transfection protocol for 30 mL culture volume

1		One day prior to transfection, prepare culture cells to be appropriate cell numbers for transfection using a 125 mL or 250 mL of Erlenmeyer shake flask. Note: Recommended seeding cell density is > 0.8 x 106 viable cells/mL.
2		[Prepare cell suspension] Just before transfection, prepare 28 mL of cell suspension with the cell densities below in a 125 mL of Erlenmeyer shake flask. ●For Expi293F™ Cells 2.7 x 106 viable cells/mL medium (7.5 x 107 viable cells/28 mL of medium) ●For FreeStyle™ 293-F Cells 1.1 x 106 viable cells/mL medium (3.0 x 107 viable cells/28 mL of medium) Note: cell viability must not be < 90 %.
3		Mix ScreenFect™ UP-293 Transfection Reagent well with a vortex mixer.
4		[Prepare a diluted transfection reagent] ●For Expi293F™ Cells Add 60 µL of ScreenFect™ UP-293 Transfection Reagent into 940 µL of ScreenFect™ UP-293 Dilution Buffer in a sterile microfuge tube and mix it using pipette strokes. ●For FreeStyle™ 293-F Cells Add 30 µL of ScreenFect™ UP-293 Transfection Reagent into 970 µL of Opti-MEM™ I Reduced Serum Medium in a sterile microfuge tube and mix it using pipette strokes.
5		[Prepare a diluted DNA solution] ●For Expi293F™ Cells Add 30 µg of plasmid DNA with a density of 1µg/µL into 970µL of ScreenFect™ UP-293 Dilution Buffer in a sterile microfuge tube and mix it using pipette strokes. ●For FreeStyle™ 293-F Cells Add 30 µg of plasmid DNA with a density of 1µg/µL into 970µL of Opti-MEM™ I Reduced Serum Medium in a sterile microfuge tube and mix it using pipette strokes.
6		[Prepare a lipoplex solution] Mix the diluted transfection reagent and the diluted DNA solution with rapid pipette strokes and spin down it. Incubate it for 5-20 minutes at room temperature. Note: Do not vortex the mixed solution.
7		Add the 2 mL of the lipoplex solution into the cell suspension in the Erlenmeyer Shaker Flask with slowly swirling. Note: The final volume should be approximately 30mL in the flask.
8		Incubate the cells at 37°C (8% CO2), rotating at 125 rpm on an orbital shaker for 16 hours.
9		Add 150 µL of ScreenFect™ UP-293 Booster to the cells.
10		Continue to incubate the cells at 37°C (8% CO2), rotating at 125 rpm on an orbital shaker. Collect cells or culture supernatants containing target proteins in 48 - 120 hours after transfection. Note: The incubation period required for successful transfection depends on the target protein and cell viability.

Expi293F™, Freestyle™ and Opti-MEM™ are registered trademarks of Thermo Fisher Scientific.

References

1. Diefenbacher, Markus E., et al. "The LIM Domain Protein nTRIP6 Recruits the Mediator Complex to AP-1-Regulated Promoters." PLoS ONE 9.5 (2014): e97549.
2. Freise, Christian, and Uwe Querfeld. "Inhibition of vascular calcification by block of intermediate conductance calcium-activated potassium channels with TRAM-34." Pharmacological Research (2014).
3. Hagiwara, Akane, et al. "Luteinizing Hormone-Induced Expression of Ptger4b, a Prostaglandin E2 Receptor Indispensable for Ovulation of the Medaka Oryzias latipes, Is Regulated by a Genomic Mechanism Involving Nuclear Progestin Receptor."
4. Peng, Yanyan, Ruidan Xu, and Xiaofeng Zheng. "HSCARG Negatively Regulates the Cellular Antiviral RIG-I Like Receptor Signaling Pathway by Inhibiting TRAF3 Ubiquitination via Recruiting OTUB1." PLoS pathogens 10.4 (2014): e1004041. (3)
5. Wakimoto, Hiroaki, et al. "Targetable signaling pathway mutations are associated with malignant phenotype in IDH-mutant gliomas." Clinical Cancer Research (2014). (2)
6. Fischer, Simon, et al. "Breaking limitations of complex culture media: Functional non-viral miRNA delivery into pharmaceutical production cell lines." Journal of biotechnology 168.4 (2013): 589-600.
7. Bai, Dongmei, et al. "Regulation of the HDM2-p53 pathway by ribosomal protein L6 in response to ribosomal stress." Nucleic acids research 42.3 (2014): 1799-1811.
8. Liu, Xing, et al. "Isocitrate dehydrogenase 2 mutation is a frequent event in osteosarcoma detected by a multi‐specific monoclonal antibody MsMab‐1." Cancer medicine 2.6 (2013): 803-814.

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