ScreenFect(TM)A plus

Outline of ScreenFect™ A plus Protocol

The 1-Step Protocol is 24 hours faster than 2-Step's.

Comparison of transfection method between 1-Step and 2-Step

➡Effective transfection for difficult-to-transfect cells!

➡Effective transfection for difficult-to-transfect cells!

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 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)

Experiment of transfecting GFP fusion gene to hiPSC (201B7)

The experiment of transfection of GFP fusion gene to hiPSC (201B7) was done by reverse transfection method (1-step) and the expression level of transfected gene was compared by a fluorescence microscope.
Reverse transfection method is the most efficient in transfection into hiPSC(201B7). It was demonstrated that the transfection efficiency with ScreenFect™A plus is equal or superior to competitor’s product in both StemSure hiPSC medium and mTeSR™1 medium.

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

In comparison with competitor’s product in the hiPSC(201B7)

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  Transfection conditions of ScreenFect™ A plus
  Cell Number	5×105 cells/well
  Amount of Plasmid DNA	4 µg/assay
  Ratio	pDNA (µg):ScreenFect™ A plus reagent(µL) = 1:0.5
  Well Format	12 well plates
  Note	SFA plus reagent and pDNA were diluted by Opti-MEM.

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  Transfection conditions of competitor's product
  Cell Number	5×105 cells/well
  Amount of Plasmid DNA	2 µg/assay
  Ratio	pDNA (µg):ScreenFect™A plus reagent(µL) = 1:2
  Well Format	12 well plates

Experiment of tranasfecting PIK3CB siRNA to HeLa cells (adherent cells)

The experiment of transfection of PIK3CB siRNA to HeLa cells was done by both methods of the reverse transfection (1-Step) and the forward transfection (2-Step). The expression quantity of PIK3CB mRNA was measured by real-time quantitative PCR.
Compare the knockdown efficiency from the quantified result against the competitor, ScreenFect™A plus showed knockdown efficiency is equal or superior to the competitor’s product.

Comparisons the pereformance of HeLa cells (adherent cells)

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  Cell Number	1×105 cells/well
  Amount of siRNA	5 pmol/assay
  Amount of Transfection Reagent	ScreenFect™ A plus reagent = 0.5 ~ 1.5 μL Company A's product = 1.5 μL
  Well Format	24 well plates
  Detection Time	48 hours after the transfection

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  Cell Number	0.5×105 cells/well
  Amount of siRNA	5 pmol/assay
  Amount of Transfection Reagent	ScreenFect™ A plus reagent = 0.5 ~ 1.5 μL Company A's product = 1.5 μL
  Well Format	24 well plates
  Detection Time	48 hours after the transfection

List of Cells transfected by ScreenFect™ A or A plus

FUJIFILM Wako Pure Chemical Corporation is updating constantly the ScreenFect™ Databese.
If you are not sure whether it is applicable to cells other than listed above, feel free to ask us.

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.