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DNA and siRNA transfection reagent

ScreenFect™A plus

A new liposome developed by click chemistry!!

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

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

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

ScreenFect™ Sample
Ordering form.

Please fill out the answers as much as possible. We will give the sample pack of ScreenFect™.

ScreenFect™
Sample Ordering form

Questionnaire after the
ScreenFect™ has been used.

Please fill out the answers as much as possible. We will give a small gift to the people who complete the questionnaire.

Questionnaire after
the ScreenFect™ has been used.

ScreenFect™A plus Transfection performance

MCF-7 (Human breast cancer (adherent)

* 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!

* 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!

* 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!

Application data

1. 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)

images

[Cell number] 3×105cells/well
[Amount of plasmid DNA] 1µg/assay
[Ratio] pDNA (µg):ScreenFect™A plus reagent(µL)=1:3
[well format] 24 well plates

images

[Cell number] 1.5×105cells/well
[Amount of plasmid DNA] 1µg/assay
[Ratio] pDNA (µg):ScreenFect™A plus reagent(µL)=1:3
[well format] 24 well plates


2. 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


3. 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)

 

images

Transfection conditions of ScreenFect™A plus
[Cell number] 5×105cells/well
[Amount of plasmid DNA] 4µg/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.

images

Transfection conditions of competitor's product
[Cell number] 5×105cells/well
[Amount of plasmid DNA] 2µg/assay
[Ratio] pDNA (µg):ScreenFect™A plus reagent(µL)= 1:2
[well format] 12 well plates


 

images

images

Transfection conditions of ScreenFect™A plus and competitor's product
 [Cell number] 5×105cells/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.


4. 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)

SFA_plus_PIK3CB_01_eng

[Cell number] 1×105cells/well
[Amount of siRNA] 5pmol/assay
[Amount of transfection reagent]
 ScreenFect™A plus reagent = 0.5 ~ 1.5 μl
 Company A's product = 1.5μl
[well format] 24 well plates
[Detection time] 48 hours after the transfection

SFA_plus_PIK3CB_02_eng

[Cell number] 0.5×105cells/well
[Amount of siRNA] 5pmol/assay
[Amount of transfection reagent]
 ScreenFect™A plus reagent = 0.5 ~ 1.5 μl
 Company A's product = 1.5μl
[well format] 24 well plates
[Detection time] 48 hours after the transfection


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

Outline of ScreenFect™A plus Protocol

images

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

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.

For details, please refer to the detailed information from the URL (http://screenfect.jp) or QR code at left. We also have free sample for evaluation, so please request from sample order form.

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 / 25μL 0.5.~ 1.5 μL/25 μL
12 wells 4 cm2 1,000 μL 100 μL 10 pmol / 50μL 1.0 ~ 3 μL/50 μL
6 wells 10 cm2 2,000 μL 250 μL 25 pmol / 125μL 2.5 μL/ 125 μL

Note: Although ScreenFect™A plus works well for siRNA transfection, we highly recommend the use of our specialized reagent ScreenFect™siRNA. Please visit the page.

List of Cells transfected by ScreenFect™A or A plus

For further information, refer to the ScreenFect™ Databese.

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

Wako Pure Chemical Industries, Ltd. 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.

ScreenFect™A plus product information

Now on sales campaign! We will offer special price during the campaign.
[Term of campaign : November 11, 2016 to February 28, 2017 ] For further information, inquires to our distributors.

Code No. Product Name Transfection Reagent Dilution Bufferfor Price (JPY)
293-77101
refrigerator
ScreenFect™A plus
0.2ml 10ml 9,000
299-77103 1ml 50ml 35,000
297-77104 1ml×5 5×50ml 140,000

Please click a product code to check the availability on our Siyaku com.

  • icon ∙∙∙ Store at 2 to 10°C
  • icon ∙∙∙ Store at −20°C
  • icon ∙∙∙ Store at −80°C
  • Store at room temperature if not specified.
  • icon ∙∙∙ Specified poisonous substance
  • iconicon ∙∙∙ Poisonous substance
  • iconiconicon ∙∙∙ Deleterious substance
  • icon ∙∙∙ Class 1 Specified Chemical Substance by the Evaluation of Chemical Substances and Regulation of Their Manufacture, etc. (CSCL)
  • icon ∙∙∙ Class 2 Specified Chemical Substance by the CSCL
  • icon ∙∙∙ Poisonous substance
  • icon ∙∙∙ Deleterious substance
  • icon ∙∙∙ Class 1 designated substance by the Act on the Prohibition of Chemical Weapons and the Regulation of Specific Chemicals,
  • icon ∙∙∙ Class 2 designated substance by the same act
  • icon ∙∙∙ Psychotropic
  • icon ∙∙∙ Raw material of specified narcotics and psychotropics
  • icon ∙∙∙ Cartagena Protocol

* The details given show the information available as of October, 2016 . Please refer to siyaku.com (http://www.siyaku.com/) for any laws and up-to-date information other than those mentioned above.

The reagent given is intended only for the purposes of testing and research and cannot be used as a “drug”, “food” or “household product”.
Suggested delivery prices show the “price of the product alone” not including consumption tax.
Suggested delivery prices are the prices at the time of preparation of this article. Please check up-to-date prices using the code No. in “Product search”.

ScreenFect™ 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.

  Others

Wako Pure Chemical Industries, Ltd

Web Site

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Head Office 1-2 Doshomachi 3-Chome, Chuo-ku, Osaka 540-8605,
Telephone: 06-6203-3741 (switchboard)
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Web Site

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Head Office Richmond, VA, Telephone: 1-804-714-1920, Facsimile: 1-804-271-7791
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Web Site
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