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产品介绍:
利用我们的创新和专有的脂质体共轭合成技术,LipoJet™转染试剂盒,运用了新型的含氟阳离子脂质体配方,同市场上的其他脂质体转染试剂有着显著 的区别,其特点是转染效率高、毒性小。LipoJet™转染试剂盒是最强大的基因 转导工具,适于几乎有所有体外基因转导的应用,包括质粒DNA、siRNA和shRNA转染和DNA/siRNA共转等。
试剂盒内容:
- LipoJet™ 试剂,1.0ml,足够用1000次DNA的转染(24孔板中0.5 ug DNA/孔)足够用1000次siRNA转染(24孔板中10 nM siRNA/孔)。
- LipoJet™转染缓冲液(5X ),按照最大化的转染效率配制,8.0ml浓缩液就可以制成40ml的工作液体。
产品特点:
- 适用于广泛的哺乳动物细胞
- 极高效率地转染DNA,siRNA和DNA/siRNA共转染
- 10 nM siRNA能达到95%的沉默效应
- 无血清和抗生素干扰
- 简单易用的操作步骤
- 适用于高通量的应用
- 细胞毒性最低
储存条件 :
40C储存。若储藏合适,产品的稳定性能保持12个月以上。
LipoJet™体外转染试剂具有极广谱的转染活性。细胞类型
DNA转染效率
siRNA转染效率
293, 293T
3T3, NIH 3T3
3T3-L1
A549
B16-F10
BNLCL.2
C2C12
C3H/10T1/2
Caco-2
Rat primary cardiomyocytes
CHO
COS1
COS7
Human bladder carcinomal cell
mES
iMEF
HBEC
HCT116
Hela
HepG2
HT-29
HuH-7
LNCaP
MC3T3-E1
MCF10A
MCF7
MDA-MB-231
MDCK
MEF
Human primary melanocytes
Human primary melanoma
Human immortalized microglial
mIMCD-3
MRC-5
N2A
PC3
RAW 264.7
RPE
SH-SY-5Y
SK-OV-3
U-2OS
VERO85%
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30%
80%
75%
55%
80%
60%
20%
10%
80%
70%
75%
80%
50%
55%
45%
75%
80%
60%
45%
45%
45%
50%
40%
50%
50%
20%
50%
50%
50%
50%
50%
50%
68%
70%
45%
30%
75%
70%
80%
40%-
90% at 20 nM
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90% at 20 nM
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90% at 40 nM
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80 % at 30 nM
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85% at 30 nM
85% at 30 nM
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90% at 30 nM
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以下图例表明LipoJet™体外转染试剂盒出色的DNA和siRNA转染效率 。
A efficiency comparison of LipoJet™ reagent vs. brand name products to transfect Hela, Cos-7, NIH-3T3, CHO and HEK293 (left panel) and Cos-7 (right panel). pEGFP-N3 cDNA (0.5 µg/well of 24-well plate) was transfected into different mammalian cells per the standard transfection protocols in presence of serum (10% FBS) and antibiotics as recommended by manufacturers. The transfection efficiency were analyzed via FACS 48 hours post transfection.
A toxicity comparison of LipoJet™ reagent vs. brand name products to transfect Hela cells. pEGFP-N3 cDNA (1.0 µg/well of 24-well plate) was transfected to Hela cells per the standard transfection protocols in presence of serum (10% FBS) and antibiotics as recommended by manufacturers. The MTT assay (right panel) and phase contrast imaging (left panel) were used to analyze the cell viability 48 hours post transfection.
A comparison of LipoJet™ reagent vs. Lipofectamine 2000 (L2K) and PolyJet™ transfection reagent on HEK293T cell. pEGFP-N3 cDNA (0.125 µg/well, 0.25 µg/well and 0.5 µg/per well of 24-well plate) was transfected into 293T cells using the standard transfection protocols in presence of serum (10% FBS) with LipoJet™ (upper panel at LipoJet™/DNA (µl/µg) ratio=2), L2K (middle panel at L2K/DNA (µl/µg) ratio=3) and PolyJet™ (lower panel at at PolyJet™/DNA (µl/µg) ratio=3) respectively. The cells were visualized by Nikon Eclipse Fluorescence microscope 48 hours post transfection.
A comparison of LipoJet™ reagent vs. Lipofectamine 2000 (L2K) and PolyJet™ transfection reagent on Hela cell. pEGFP-N3 cDNA (0.125 µg/well, 0.25 µg/well and 0.5 µg/per well of 24-well plate) was transfected into Hela cells using the standard transfection protocols in presence of serum (10% FBS) with LipoJet™ (upper panel at LipoJet™/DNA (µl/µg) ratio=2), L2K (middle panel at L2K/DNA (µl/µg) ratio=3) and PolyJet™ (lower panel at at PolyJet™/DNA (µl/µg) ratio=3) respectively. The cells were visualized by Nikon Eclipse Fluorescence microscope 48 hours post transfection.
LipoJet™试剂介导的DNA/siRNA共转染表现出色的基因沉默效应。分别由LipoJet™试剂转染GFP的cDNA (左图, 0.25µg/孔,24孔板)作为对照和共转染GFP的cDNA (0.25µg/孔,24孔板)/GFP的siRNA (终浓度10 nM,右图)至HEK293细胞 (上图), Hela细胞(中图)和SaoS-2细胞(下图)。转染24小时后,使用尼康Eclipse荧光显微镜检查GFP荧光。结果显示,GFP的cDNA和GFP的siRNA共转染导致GFP的表达被显著抑制。
Exceptional DNA/siRNA co-transfection efficiency on HUVEC. Co-transfection of mCherry cDNA (0.10 µg per well of 24-well plate) and FITC conjugated siRNA (final 30 nM per well of 24-well plate) to HUVEC with LipoJet™ reagent gave rise to 60% mCherry+ (phase contrast overlapped with mCherry imaging, left panel) and nearly 100% FITC-siRNA+ (phase contrast overlapped with FITC imaging, right panel) HUVEC 24 hours after transfection. The pictures were given from Dr. Pan Kong of USC as courtesy.
Excellent silencing of endogenously expressed KIF11 (also known as EG5) in HEK293 (upper panel) and Hela (lower panel) cells with LipoJet™ reagent at 10 nM EG5 siRNA. KIF11 (also known as EG5) encodes a motor protein that belongs to the kinesin-like protein family involved in chromosome positioning and bipolar spindle formation during cell mitosis. A reduction in KIF11 levels causes mitotic arrest. LipoJet™ reagent effectively delivers EG5 siRNA (final 10 nM) to HEK293 and Hela cells, leading to more than 80% of "round-up" phenotype of HEK293 and Hela cells 24h post transfection over negative control (final 10 nM with sham EG5 siRNA). The phenotype of "rounded-up" HEK293 and Hela cells were visualized 24h post transfection with a Nikon microscope.
LipoJet™转染试剂介导的基因沉默可显著地抑制HEK293细胞(上图)和Hela(下图)内源型EG5的表达。KIF11 (也称为EG5)编码了一个启动蛋白,这启动蛋白属于驱动蛋白家族,参与染色体定位和细胞有丝分裂时两端的纺锤体的形成。EG5水平降低会阻碍有丝分裂。LipoJet™试剂能有效的将EG5 siRNA(终浓度10 nM)导入到HEK293和Hela细胞中。转染24小时后,参照阴性对照(终浓度是10 nM+假EG5 siRNA),LipoJet™介导的EG5 siRNA的导入显著地抑制了细胞有丝分裂,导致了大于80%的细胞出现圆顶表型。