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New Liposome developed by click chemistry
ScreenFect™ A
< DNA & siRNA Transfection Reagent >

ScreenFect™ A is a transfection reagent composed of new cationic lipids selected by screening experiments *1 by click chemistry. It is applicable to cell lines from various eukaryotic species and can be directly added to medium containing antibiotic and serum. ScreenFect™ A works with DNA and siRNA and can show highly efficient transfection in common cell lines (HeLa, HepG2, MDCK, Cos‐7, etc.), stem cells (mouse embryonic stem cells, etc.), blood cells (macrophage, THP‐1, RAW264.7, etc.), microglia, primary culture cells.
Because of the low cell toxicity, medium change after transfection is not required. Furthermore, this kit does not contain any poisonous and deleterious components.
*1: Biomaterials. Nov., 33 (32): 8160‐6. 2012
LINK to ScreenFect Database
ScreenFect was advertised in nature
on Nov. 27, 2104. ⇒ zoom


1. Features

  1. Highly efficient transfection & Low cell toxicity
  2. Applicable to both DNA & siRNA
  3. Requires no medium change after transfection
  4. Serum Compatible
2. Product Comparison

GFP plasmid was successfully transfected into HEK 293 cell using ScreenFect™ A. The transfection efficiency is the same or more than that of competitor’s products commercially available. (using 96 well plate, GFP‐expressing plasmid: 75 ng/well)



3. Transfection Efficiency of Liposome Library

GFP‐expressing plasmid was transfected into HEK 293T cells using a library of newly developed cationic liposome synthesized by click chemistry. Company A’s conventional product is shown as positive control. ScreenFect&trade: A showed the best efficiency at minimal toxicity.


4. ScreenFect™ A Protocol

♣ DNA Transfection (96 wells)
DNA transfection (per well)
Plate Size Surface Area Medium Volume Total Volume
SF-DNA complex
DNA/Dilution Buffer ScreenFect™ A/Dilution Buffer
96 wells
0.3 cm2
~ 80 μL
16 μL
~ 75 ng / 8 μL
0.2 ~ 0.4 μL / 8 μL
24 wells
2 cm2
~ 350 μL
80 μL
~ 300 ng / 40 μL
1 ~ 2 μL / 40 μL
12 wells
4 cm2
~ 700 μL
140 μL
~ 600 ng / 70 μL
2 ~ 4 μL / 70 μL
6 wells
10 cm2
~ 1,250 μL
240 μL
~ 1000 ng / 120 μL
4 ~ 6 μL / 120 μL

♣ siRNA Transfection (24 wells)
DNA transfection (per well)
Plate Size Surface Area Medium Volume Total Volume
SF-siRNA complex
DNA/Dilution Buffer ScreenFect™ A/Dilution Buffer
96 wells 0.3 cm2 84 μL 16 μL 2 ~ 3 pmol / 8 μL 0.15 ~ 0.35 μL / 8 μL
24 wells 2 cm2 420 μL 80 μL 10 ~ 20 pmol / 40 μL 0.75 ~ 2 μL / 40 μL
12 wells 4 cm2 750 μL 150 μL 20 ~ 60 pmol / 75 μL 1.5 ~ 4 μL / 75 μL
6 wells 10 cm2 1,260 μL 240 μL 20 ~ 60 pmol / 120 μL 2.5 ~ 6 μL / 120 μL

Please refer to the package insert how to use ScreenFect™ A. As for the protocol for optimization on cell species, please contact us.

5. ScreenFect™ A Applications
♣ Low Cell Toxicity




GFP‐expressing plasmid was transfected into HEK 293 using ScreenFect™ A. ScreenFect™ A is superior to Company A’s product on transfection efficiency and comparable to company B’s one on low cell toxicity. (96 well plate, GFP‐expressing plasmid 75 ng/well)


♣ Transfection of Mouse Embronic Stem Cells (mESCs) with ScreenFect™ A



GFP transfected cells (left) and bright field (right). Transfection efficiencies of approx. 60% are typically obtained, with low cell toxicity with ScreenFect™ A.



♣ Examples of ScreenFect™ A mediated siRNA delivery and Resulting Protein Knock-Down





LRP6 siRNA was transfected into HEK 293 using ScreenFect™ A. Knowckdown effect is stronger using ScreenFect™ A than Company A and B. (96 well plate, 1 pmole LRP6 siRNAFinal concentration: 2 nM/well)



GAPDH siRNA was transfected into various cell lines using ScreenFect™ A. Knowdown effect is stronger than Company A’s product. (96 well plate, GAPDH siRNA Final Concentration: 3 nM/well)

6. ScreenFect™ A Transfection & Example of Cell Lines Transfected
♣ Transfection Efficiency
Cell Strain Nucleic Acid Scale Transfection Efficiency
(Time required for detection)
Indication
HEK293
DNA
6 - 96 well
70-98% (24-48h)
GFP
HEK293
DNA
6 - 96 well
70-98% (24-48h)
Luciferase
HeLa
DNA
6 - 24 well
50-70% (24-48h)
GFP
MCF7
siRNA
6 well
70-98% (48h)
Specific Gene Expression
MEF
DNA
6 well
20-50% (24-48h)
GFP
mC3T3
DNA
6 well
50-70% (24-48h)
GFP
LO2
DNA
12 well
20-50% (48h)
Antibiotic-resistant
MCF-10A
siRNA
6 - 12 well
10-20% (48h)
GFP
HepG2
DNA
6 well
50-70% (48h)
GFP
Du145
DNA
6 -12 well
10-20% (24h)
GFP
RAW264.7
DNA
6 well
20-50% (24-48h)
GFP
HL7704
miRNA
6 well
70-90% (48h)
Specific Gene Expression
H9C2
DNA
6 well
10-20% (48h)
GFP
U937
DNA
6 well
20-50% (24-48h)
GFP
HepG2
DNA
24 well
50-70% (12h)
GFP
HUVEC
DNA
24 well
10-20% (24-48h)
GFP
THP1
miRNA
6 well
60-80%
Specific Gene Expression
PLC8024
DNA
12 well
10-20% (48h)
GFP
mES
DNA
6 - 24 well
40-60% (24-48h)
GFP
HEp-2
DNA
6 - 24 well
50-70% (48h)
GFP

♣ Example of Cell Lines Successfully transfected with ScreenFect™ A Transfection Reagent
No. Cell Strain Source
1
HEK293
human embryonic kidney cells
2
HELA
cervical cancer cells
3
MCF7
breast cancer cells
4
A431
epidermoid carcinoma cells
5
MEF
mouse embryonic fibroblasts
6
mESC
mouse embryonic stem cells
7
PAC2
fibroblast zebrafish cells
8
MTPa
mammary adenocarcinoma cells
9
AB9
caudal fin of adult zebrafish
10
Ins‐1
murine insulinoma cells
11
P388D1
Macrophages
12
M3CT3‐E1
osteoblast like cell line
13
LO2
human hepatic cells
14
MCF‐10a
human mammary epithelial cells
15
HepG2
human liver carcinoma cells
16
Du14S
human prostate carcinoma cells
17
RAW264.7
murine macrophage leukemia cells
18
H9C2
rat myoblast cells
19
U937
human histiocytic leukemia cells
20
HUVEC
human umbilical vein endothelial cells
21
HL7704
human adult hepatocyte cells
22
BEL7402
human hepatocellular carcinoma cells
23
THP‐1
human acute monocytic leukemia cells
24
SK‐Hep1
human liver adenocarcinoma cells
25
786‐O
human kidney carcinoma cells
26
HKC‐8
human renal proximal tubular cells
27
NE3
human esophageal immortalization cells
28
PLC8024
human hepatoblastoma cells
29
NIH3T3
mouse embryonic fibroblast cells
30
MG6
mouse microglia cells
31
MDCK
madin‐Darby canine kidney cells
32
Cos‐7
monkey Kidney Fibroblast cells
33
HEp‐2
human epidermoid cancer cells
34
 
Primary cells form Liver tissue
Application data as of April, 2013
Wako is regularly updating the application data of ScreenFect™ A. Please contact us about the applicability of your cell strains.
7. Products List
ScreenFect™ A            <Wako Catalog No.>
293-73201 299-73203 297-73204
<Packge Size>
0.2 mL 1 mL 1 mL x 5
                      Kit Contents
Transfection Reagent
0.2 mL
1 mL
1 mL x 5
Dilution Buffer
10 mL
50 mL
50 mL x 5

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