LipoJet™转染试剂盒(Ver. II)

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。

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。

转染试剂操作步骤及其使用指南 ：
– LipoJet™试剂DNA和siRNA转染操作步骤
– 简易DNA转染操作步骤(适合熟练操作者) 
– 简易siRNA转染操作步骤(适合熟练操作者) 
– LipoJet™试剂DNA/siRNA共转染操作步骤 
– 注意事项及转染窍门 

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用户评价：

I tried the plasmid/siRNA co-transfection using the LipoJet sample in EAhy926 cell which is a HUVEC cell line. The results are pretty good, at least for the transfection efficiency. For the plasmid transfection, 48 hours later, around 60% cell are transduced. For siRNA, the efficiency is almost 100%. We are ordering more LipoJet reagent to use from now.

—–Dr. Pan Kong, University of Southern California

We got >90% efficiency with LipoJet vs. 80% with X-tremeGENE 9 on 293T cell. Definitely will order more….

—–Dr. Nuo Yang from Roswell Park Cancer Institute

Here are the results from our preliminary experiments with LipoJet. We are very satisfied with its efficiency. Unfortunately, we were out of Fugene 6/HD and were unable to test them side by side, but based on previous experience, we do believe that your product worked with better efficiency.

——Dr. Alison McKelvey from University of Pittsburgh

I am happy to say that I had great success with your transfection reagents on OKF6/TERT2 human keratinocytes. I will put together the data once I have completed my analysis. I really like the LipoJet. My cell of interest did not do very well in the GenMute although my control cells did. We want to go ahead with ordering ASAP. I also have a coupon for 10% for a PO order.

—–Dr. Michelle Simpson-Abelson from UPMC

We did indeed test them side by side with our homemade CaPhos. The results are great,  LipoJet and CalFectin both did extremely well. LipoJet being the better of the two. I will send you the results when  I get them,  I am currently out of the lab but we will be ordering more for certain. Thank you for sending those samples.

——Dr. Ahmed Hassib, Cornell University

For the LipoJet, I only compared it with Lipo LTX in Hela cells. The LipoJet gave a much higher efficiency (~35% better than Lipo LTX) 24 hours after transfection. We will order some LipoJet.

—–A beta tester from USC