SFA P-reagent

SFA P-reagent enhances transfection efficiency and expression level in each transfected cells by using ScreenFect™ together when plasmid DNA and mRNA are transfected. Additionally, cytotoxicity also decreases remarkably by adding SFA P-reagent.

SFA P-reagent will resolve the following problems.

• Enhances transfection efficiency
• Increases the expression level of transfected gene
• Decreases cytotoxicity
• Saves the amount of plasmid DNA (Ex. : Half (50%) of current condition)

Experiment of transfecting YFP fusion gene to LNCaP cell (adhesive type)

The experiment of transfection of YFP fusion gene to LNCaP cells (adhesive type) was done, and the expression level of transfected gene was compared by a fluorescence microscope.
Using both ScreenFect™A plus and SFA P-reagent as enhancer reagent, it was demonstrated that the expression efficiency is equal or superior to other competitor’s products.

*Described data in BioWindow No. 144 (published on June, 2016)

In comparison with competitor’s product in the LNCaP cell (human prostatic cancer)

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  Cell Number	3 x 105 cells/well
  Amount of Plasmid DNA	1 µg/assay
  Ratio	pDNA(µg): ScreenFect™ A plus reagent(µL) = 1:3
  Well Format	24 well plates

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  Cell Number	1.5 x 105 cells/well
  Amount of Plasmid DNA	1 µg/assay
  Ratio	pDNA(µg): ScreenFect™ A plus reagent(µL) = 1:3
  Well Format	24 well plates

Experiment of transfecting plasmid DNA to HeLa cell

The experiment of transfection of GFP fusion gene to HeLa cells (adhesive type) was done, and the expression level of transfected gene was compared by a fluorescence microscope.
Using both ScreenFect™ A plus and SFA P-reagent, it was demonstrated that the expression efficiency is equal or superior to competitor’s products.

Cell Number	1.5 x 105 cells/well
SFA P-reagent	0.5 µL/well
Ratio	Plasmid DNA(µg): ScreenFect™A plus reagent(µL) = 1 : 4
Well Format	24 well plates
Detection Time	24 hours after transfection

Experiment of transfecting EGFP_mRNA to HeLa cell (adhesive type)

The experiment of transfection of EGFP_mRNA to HeLa cells (adhesive type) was done, and the expression level of transfected gene was compared by a fluorescence microscope.
Using both ScreenFect™A plus and SFA P-reagent as enhancer reagent, it was demonstrated that the expression efficiency is equal or superior to other competitor’s products.

*Described data in BioWindow No. 144 (published on June, 2016)

In comparison with competitor’s product in the LNCaP cell (human prostatic cancer)

Cell Number	0.7×105 cells/well
Amount of mRNA	0.1µg/assay
Ratio	mRNA(µg) : ScreenFect™A plus reagent(µL) = 1 : 4
Well Format	24 well plates
Detection Time	48 hours after transfection
Time of Exposure	2 seconds

Data of cytotoxity reduction

The experiment of transfection of luciferase reporter vector to CHO-K1 was done by reverse transfection method (1-step) and compared expression level and cell viability.
The cell viability of cells was increased significantly by adding SFA P-reagent, and expression level was almost as good as that of other company.

*Described data in BioWindow No. 145 (published on September, 2016)

Cell Number	2×104 cells/well
Amount of plasmid DNA	100 ng/assay
Ratio	pDNA(µg) : ScreenFect™A plus reagent(µL) = 1 : 3 pDNA(µg) : SFA P-reagent(µL) = 1 : 2
Well Format	96 well plates

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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