简介: 根据SignaGen公司专利聚合物合成技术,即PolyJet 技术,脱氧核糖核酸体外转染试剂被指定为一种可生物降解的聚合物转染试剂,确保HEK293, COS-7, NIH-3T3,Hela,CHO及多种难以被转染的哺乳细胞等的有效且可再生转染。PolyJet试剂能在电泳期间低浓度固定脱氧核糖核酸迁移,并在5分钟内通过RT迅速形成转染复合体。试剂的一个卓越的特点是聚合物能够在转染聚合物细胞内吞(见图1)后迅速并完全降解聚合物,这样可大大降低细胞毒性。仅1.0 ml的PolyJet试剂就足够进行24个试剂盒的300到600个细胞转染或6个试剂盒的150到300个细胞转染,这样就为各种常用的和难以转染的哺乳细胞等主导产品提供一个非常充裕的选择。
Cell Lines | Efficiency (% GFP) | Cell Lines | Efficiency (% GFP) |
McArdle 7777 Hep3D SHEP 3T3-442A COS-7 CV-1 D 407 DHD Pro.b 3LL B16-F10 BAEC BHK-21 Ca Ski CaCo2 CHO HCS-2/8 HEK-293 HeLa HLMEC H-MVEC Huh-7D12 ATT20 SK-N-SH C2C12 HepG2 | 65-70% 67-76% 68-71% 35% 85-90% 60% 70% 70% 80% 85% 51% 80% 88% 60% 88% 61% 86% 88% 72% 59% 72% 46% 29% 46% 72% | SAOS-2 SN56 MC3T3-E1 Primary melanocyte K562 L929 MCF-7 MDCK Neuro2A NIH 3T3 PC12 SH-SY5Y SiHa SKOV3 Huh-7 IGROV1 DF-1, Chicken Embryonic Cell 6CSFMEo WEHI 231 A549 LNCap Prim. mouse keratinocyte Prim. human skin fibroblast Prim. human pre-adipocyte Prim. mouse embry. fibroblast | 58% 81% 80% 35% 38% 59% 68% 68% 86% 76% 50% 25% 60% 65% 70% 35% 50% 71% 26% 75% 75 29% 50% 32% 30% |
针对中国仓鼠卵巢(CHO)细胞,PolyJet转染效率同lipofectamine Plus试剂[由Life Technologies公司(BRL)生产的商品化细胞转染试剂,是合成的阳离子脂类DOSPA和DOPE的3:1(w/w)混合物]进行比较。标记HA的Beta微管蛋白DNA分别加入PolyJet试剂(左图)和lipofectamine Plus试剂 (右图)被输送进仓鼠卵(胞。 HA标记的FITC共轭抗体被用于提取HA-beta微管蛋白 (绿色部分),而DM1a抗体被用于检测伴随罗丹明共轭二次抗体的内源阿尔法微管蛋白 (红色部分)。上述图片由得克萨斯大学休斯敦医学院的Shang Yin博士友情提供提供。 研究(通过HEK293FT细胞试验,PolyJet试剂的同主导试剂产品Lipofectamine 2000相比后所展示的转染效率)。 HEK-293FT细胞通过分别输入与GFP传染媒介(pEGFP-N3)结合的PolyJet 试剂(左图) 和Lipofectamine 2000(右图)被转染。该细胞在转染后24小时可通过尼康Eclipse荧光显微镜进行观察。 比较研究(通过HepG2细胞试验,PolyJet试剂的同主导试剂产品Lipofectamine 2000相比后所展示的转染效率)。HepG2细胞通过分别输入与GFP传染媒介(pEGFP-N3)结合的PolyJet 试剂(左图) 和Lipofectamine 2000(右图)被转染。该细胞在转染后24小时可通过尼康Eclipse荧光显微镜进行观察。 Transfection efficiency comparison of PolyJet™ vs. Fugene HD on MDCK cells. A plain GFP DNA was transduced into MDCK cells with PolyJet™ (left panel) and Fugene HD (right panel) reagents respectively per manufacturers protocols. GFP and DAPI staining were visualized under fluorescence microscopy 48 hours post tansfection. The above comparison data and pictures were completed and provided by Dr. Ge Zhou of NYU Medical Center as courtesy 比较研究(通过MDCK细胞试验,PolyJet试剂的同主导试剂产品Lipofectamine 2000相比后所展示的转染效率)。MDCK属于极难转染的细胞。根据专有“削减细胞转染”协议,PolyJet 试剂(左图) 放弃了70%的GFP阳性细胞,相比之下,Lipofectamine 2000(右图)的效率仅约为5%。MDCK细胞通过分别输入与GFP传染媒介(pEGFP-N3)结合的PolyJet 试剂(左图) 和Lipofectamine 2000(右图)被转染。该细胞在转染后36小时可通过尼康Eclipse荧光显微镜进行观察。 比较显示转染效率的PolyJet ™试剂与领先的产品, Fugene高清的LNCap细胞。 LNCap细胞转染绿色荧光蛋白载体( pEGFP - N3型)的PolyJet ™ (左图)和Fugene高清(右)分别。转染24小时后,尼康的Eclipse荧光显微镜显示的差别图。 Neuro2A细胞通过输入与pEGFP-C1质粒相结合的PolyJet体外脱氧核糖核酸转染试剂被转染。Neuro2A细胞由转染后24小时可通过带DIC阶段显像(左)和FITC显像(右)的尼康Eclipse荧光显微镜进行观察。 . PolyJet体外脱氧核糖核酸体外转染试剂运用于主要鼠科皮肤纤维细胞上的细胞毒性与L2K进行比较。主要鼠科的纤维细胞与上述转染试剂/pEGFP-C1 (脱氧核糖核酸)的复合体在完全不含血清的DMEM高葡萄糖媒介中和含有血清的媒介中分别培养达4个小时后进行观察。该细胞由转染后24小时可通过带DIC阶段显像的尼康Eclipse荧光显微镜进行观察。技术资料及数据清单 数据清单及协议草案 - General Protocol for Transfecting Mammalian Cells - Advanced Protocol for Transfecting Hard-to-Transfect Mammalian Cells - Protocol for Lentivirus Production 用户推荐: PolyJet Transfection Reagent worked equally as well as lipofectamine 2000, with little evidence of cell death on 293, PC-3 and 22RV1 cells. I will defiantly consider switching over.-----Tiffany Wallace, Ph.D., June 9, 2009, NCI / NIHI only did side by side with the testing sample (PolyJet) and Lipo2000 with GFP transfection on COS-7 cells. The result was very good. PolyJet was even better than L2K.------Feng Qiao, Ph.D., Jun 12, 2009, NEI / NIHI tested the sample of PolyJet on my NIH-3T3 mouse fibroblasts this weekend. The results were much better than Lipofecatmine LTX. Im attaching a powerpoint slide with my results (I did not quantify the % transfection efficiency, but the pictures get the point across). I found that the protocol for difficult-to-transfect cell lines worked better than the standard protocol.-----Stephanie Murphy, Ph.D., June 29, 2009, Dartmouth CollegeI had chance to try your product finally. It was great success. I used HeLa cells and got 10% transfection efficiency (<0.1% for Lipofectamine). Thank you! I was wondering if I also try GenJet™ Plus DNA In Vitro Transfection Reagent? According to your website, the reagent works better than regular PolyJet. -------Yumi Uetake, Ph.D., August11, 2009, UMASSI tested PolyJet and it looks great on MDCK. We placed order. Thank you!-------Ge Zhou, Ph.D., August 12, 2009, NYUWe are happy to provide feedback. PolyJet worked very well for us in HepG2 cells, we got approximately 80% efficiency with pMAX GFP plasmid, by following the conditions in your suggested protocol. We ran a comparison with Lipofectamine, which only showed approximately 20-30% transfection efficiency. We are planning experiments and will be ordering more soon.-------Emily Mcallister, September 3rd, 2009, PBRC······ ······