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ScreenFect™A plus

for Genetic Research
Manufacturer :
FUJIFILM Wako Pure Chemical Corporation
Storage Condition :
Keep at 2-10 degrees C.
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Distributor
293-77101
Barcode No
4548995045218
0.2mL
List Price
99.00 USD

In stock in Japan

Distributor
299-77103
Barcode No
4548995045225
1mL
List Price
385.00 USD

In stock in Japan

Distributor
297-77104
Barcode No
4548995045232
1mL x5
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Document

SDS
Product Specification Sheet
Spectral Data
Certificate of Analysis
Calibration Certificate

Overview

ScreenFect™A plus is a transfection reagent composed of a new cationic loposome screened*1 by click chemistry.
It can be used with various eukaryote-derive cells and can be directly added to medium containing antibodies or serum.
DNA and siRNA can be transfected into general experimental cell strains (HeLa, HepG2, MECK, etc.), stem cells (mouse ES cells, etc.), blood cells (macrophages, THP-1, RAW264, 7, etc.), microglia, primary (initial subculture) cells, and insect cells. Medium replacement after transfection is not required due to low cytotoxicity. The constiuent reagents do not contain any poisonous or deleterious substance.

  • Broad Range Transfection
  • One-step Transfection
  • High Transfection Efficiency and Low Toxicity
  • Low Running Cost

*1: Biomaterials. Nov., 33 (32): 8160‐6. 2012

Outline of ScreenFect™ A plus Protocol

02392830_img21.gif

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

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

Product ScreenFect™ A plus Company A's new product
(+ reagent)
Company A's product
Recommended Protocol 1-Step 2-Step
Experiment term 2 Days 3 Days
DNA quantity Low High
Adjustment of cell numbers Flexible
(cell preparation just before transfection)
Inflexible
(depending on cell pre-culture condition)
Detachment by trypsin Required
(just before transfection)
Required
(before cell pre-culture)
Convenience for
HTS cell-base assay
+++++ +
Medium change Depending on cell lines

Applications

ScreenFect™A plus Transfection performance

MCF-7 Human breast cancer (adherent)

  • 02392830_img01.gif
  • 02392830_img02.gif

02392830_img03.gif

*MCF-7 Human breast cancer (adherent)
*Protocol Company A’s new product (+ reagent) → 2-Step
ScreenFect™A plus1 Step


➡Effective transfection for difficult-to-transfect cells!

  • 02392830_img04.gif
  • 02392830_img05.gif

02392830_img06.gif

*MDCK Madin-Darby canine kidney (adherent)
*Protocol Company A's new product (+ reagent) → 2-Step
ScreenFect™A plus1 Step


➡Effective transfection for difficult-to-transfect cells!

  • 02392830_img07.gif
  • 02392830_img08.gif

02392830_img09.gif

*K562 Human erythromyeloblastoid leukemia (suspension)
*Protocol Company A's product & the new product (+ reagent) → 2-Step
ScreenFect™A plus1 Step


➡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)

  • 02392830_img10.jpg
    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
  • 02392830_img11.jpg
    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)

02392830_img12.jpg
Cell Number 0.7×105 cells/well
Amount of mRNA 0.1 µg/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

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)

02392830_img14.jpg
  • 02392830_img15.jpg

    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.
  • 02392830_img13.jpg

    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
02392830_img16.jpg
  • 02392830_img17.jpg
  • 02392830_img18.jpg
Transfection conditions of ScreenFect™ A plus and competitor's product
Cell Number 5×105 cells/well
Amount of Plasmid DNA 1µg/assay
Ratio pDNA (µg):ScreenFect™ A plus reagent(µL) = 1:2
Well Format 12 well plates
Note SFA plus reagent and pDNA were diluted by Opti-MEM.

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)

  • 02392830_img19.png

    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
  • 02392830_img20.png

    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

More Information

ScreenFect™ A plus Transfection Condition

DNA transfection (/well)
Plate size Surface area Medium volume Total volume
SF-DNA complex
DNA
/Dilution Buffer
ScreenFect™A plus
/Dilution Buffer
96 wells 0.3 cm2 100 μL 10 μL ~ 100 ng / 10 μL ~ 0.4 μL/10 μL
24 wells 2 cm2 500 μL 50 μL ~ 500 ng / 40 μL ~ 2.0 μL/40 μL
12 wells 4 cm2 1,000 μL 100 μL ~ 1,000 ng / 70 μL ~ 4.0 μL/70 μL
6 wells 10 cm2 2,000 μL 250 μL ~ 2,500 ng / 120 μL ~ 10 μL/120 μL

Note: For large scale transfections it helps to split sample between several tubes. For example, take 5-6 samples of the 6-well transfection volumes for 10 cm dish transfection.

ScreenFect™ A plus recommended protocol

*Quick protocols are listed in the instructions attached to the actual product. Please access from the following URL or QR code.

The one-step protocol for ScreenFect™A plus transfection is described in the upper part. The optimal DNA-to-reagent-ratio for efficient cell transfection stays constant at around 1:3 to 1:4 (DNA 100 ng:ScreenFect™A plus reagent 0.1 μL = 1:1) and we recommend splitting cells when they reach 60% ~ 80% confluence to avoid contact inhibition of cell proliferation.

siRNA transfection (/well)
Plate size Surface area Medium volume Total volume
SF-DNA complex
siRNA
/Dilution Buffer
ScreenFect™A plus
/Dilution Buffer
96 wells 0.3 cm2 100 μL 10 μL 1 pmol /5 μL 0.1 ~ 0.3 μL/5 μL
24 wells 2 cm2 500 μL 50 μL 5 pmol / 25 μL 0.5.~ 1.5 μL/25 μL
12 wells 4 cm2 1,000 μL 100 μL 10 pmol / 50 μL 1.0 ~ 3 μL/50 μL
6 wells 10 cm2 2,000 μL 250 μL 25 pmol / 125 μL 2.5 μL/ 125 μL

List of Cells transfected by ScreenFect™ A or A plus

No. Name of cell
1 143BTK
2 786-O
3 A2058
4 A375
5 A549
6 B16
7 B16F10
8 Ba/F3-CH1
9 BEAS-2B
10 BEL-7402
11 BT549
12 C2C12
13 Cell line from killifish
14 CHO-K1
15 COS-7
16 DB lymphoma
17 DC 2.4
18 Du145
19 EL4
20 Endothelium cell
21 EPC(carp)
22 GP2-293
23 H9C2
24 HCT116
25 HEK293
26 HEK293 TN
27 HEK293A
28 HEK293F
29 HEK293FT
30 HEK293T
31 HeLa
32 HeLa S3
33 HEp-2
34 HepG2
35 hiPSC
36 HK2
37 HKC
38 HL7704
39 HuH-7
40 HUVEC
41 Ins-1
42 L428
No. Name of cell
43 LLC-MK2
44 LO2
45 LoVo
46 MC3T3
47 MC3T3-E1
48 MCF-10
49 MCF-10A
50 MCF-7
51 MDCK
52 MEF
53 mES
54 mHSC
55 Microglia
56 MLEC
57 MS-1
58 Myeloid dendritic cell (MDC)
59 NB1RGB
60 NCI-H1703
61 NE3
62 NIH 3T3
63 NK92
64 OLHNI-2
65 Mouse overy cell
66 PC12
67 Plat-E
68 PLC8024
69 Primary Fibroblast
70 RAW264.7
71 SH-SY5Y
72 SK-Hep1
73 SKOV3
74 T98G
75 TE-13
76 THP-1
77 U-251 MG
78 U2OS
79 U937
80 Vero
81 Drosophira ovary somatic cell
82 HT1080
83 RH7777
84 HaCaT

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.

Overview / Applications

Outline This product is for research use only. Do not administer it to human.

ScreenFect™A plus is a transfection reagent consisting of a new cationic liposome screened by click chemistry.
It can be used with various eukaryote-derived cells and can be added directly to mediums containing antibiotics or serum. In addition, medium replacement after transfection is not required due to low cytotoxicity. And the constituent reagents do not contain any poisonous or deleterious substance. The optimization range (DNA : reagent ratio) of lipoplex was more flexible than ScreenFect™A, thus ScreenFect™A plus is improved in point of transfection efficiency compared with ScreenFect™A.

Property

Manufacturer Information

Alias

  • Transfection Reagent
    Transfection Reagent
    ScreenFectTMA plus

Product content may differ from the actual image due to minor specification changes etc.

If the revision of product standards and packaging standards has been made, there is a case where the actual product specifications and images are different.

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