Cell Transfection Reagents
| Catalog# | size | Price | Quantity | |
|---|---|---|---|---|
| 31-00110 | 1ml (250 Tfx) | $180.00 | ||
| 31-00120 | 1ml (250 Tfx) | $180.00 | ||
| 31-00130 | 1ml (250 Tfx) | $220.00 |
Epoch Life Science"s GenCarrier TM transfection reagents apply a proprietary Lipoplex-enhancing mechanism to obtain the best transfection outcome - highest transgene expression and lowest cytotoxicity.
Epoch Life Science is determined to maintain the highest standards and deliver the highestquality products and services. Every batch is strictly tested to meet the requirementsof cell-culture grade bioproducts.
GenCarrier-1TM DNA transfection reagent - works in serum containing media
GenCarrier-1TM has been used successfully to transfect DNA into HeLa, CHO-K1, COS, LNCaP,NIH3T3, U937, SF-9, S2 (Schneider - Drosophila), 293, T-24, C2C12, PANC-1, primary human keratinocytes,primary aortic smooth muscle, primary rabbit myoblasts, human bone marrow endothelial cells (HBMEC),and other cell lines. No significant toxicity was observed at working concentrations on the cell lines tested.Enough reagent is provided for approximately 250 transfections in 35-mm plates per package.
PC12 cells transfected withpCMV-GFP using GenCarrier-1(upper 2 pictures) and lipofectamine 2000 (lower 2 pictures)
GenCarrier-2TM DNA transfection reagent -works in primary cell culture GenCarrier-2TM has been used successfully to transfect DNA into COS, HepG2, PC12, NIH3T3, Jurkat, PC-12, HUVEC, primary human pancreatic islet cells, primary rat and hamsterhepatocytes, primary rat cardiomyocytes, primary male rat germ cells, primary chick retinal neurons,primary rat fibroblasts, primary human keratinocytes, and tracheobroncial cells. No significant toxicitywas observed at working concentrations on the cell lines tested.Enough reagent is provided for approximately 250 transfections in 35-mm plates per package.

Human primary pancreatic cells transfected with pCMV-bGal using GenCarrier-2 showed >50% transfection efficiency
RNAiCarrierTM siRNA transfection reagent
RNAiCarrierTM has been used successfully to transfect siRNA into HEK293, HuH-7 (human hepatoma), B16 melanoma, CHO-K1, COS-1, COS-7, NIH3T3, SF-9, S2, primary human keratinocytes, primary rat aortic smooth muscle (WKY3M), and other cell lines. No significant toxicity was observed at working concentrations used on the cell lines tested.
Enough reagent is provided for approximately 250 transfections in 35-mm plates per package.
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“转化”是指含外源基因的重组质粒(载体)将外源基因直接导入原核细胞(如细菌);
“转导”指通过重组病毒载体将外源基因导入真核细胞或原核细胞;
“转染”指重组质粒载体或游离核苷酸在脂质体等介导下进入真核细胞;
“感染”在基因转移实验中强调重组病毒载体入侵受体细胞的过程。
在使用这4 个名词时, 应仔细分析基因转移实验的四要素——转移物、载体、介导方法、受体细胞类型,而正确区 分载体和受体细胞类型是辨析的关键点。当载体是重组质粒时,如受体细胞是原核细胞应使 用“转化”,如受体细胞是真核细胞则使用“转染”;当载体是重组病毒时,如强调转移物进 入受体细胞应使用“转导”,如强调重组病毒载体进入受体细胞的过程则使用“感染”。
不建议你用MDCK细胞,非常难转染。
大家都是用什么方法挑选细胞单克隆的
单克隆:单克隆是指‘子代来源于一个母体.
细胞培养:细胞的大规模克隆.细胞培养,既包括微生物细胞的培养,细胞培养技术可以由一个细胞经过大量培养成为简单的单细胞或极少分化的多细胞.
单克隆一般常指动植物细胞的克隆,细胞培养一般是指动物、微生物等细胞的细胞克隆.
二者没有什么明显区别.单克隆在单克隆抗体制备中比较常见.其实是对骨髓瘤细胞的细胞培养.
脂质体是磷脂分散在水中时形成的脂质双分子层,又称为人工生物膜。
阳离子脂质体表面带正电荷,能与核酸的磷酸根通过静电作用将DNA分子包裹入内,形成DNA一脂复合体,也能被表面带负电荷的细胞膜吸附,再通过膜的融合或细胞的内吞作用,偶尔也通过直接渗透作用,DNA传递进入细胞,形成包涵体或进入溶酶体 其中一小部分DNA能从包涵体内释放,并进入细胞质中,再进一步进入核内转录、表达。
我刚开始做转染,悬浮细胞,分别做过表达和敲减,看了很多文献,大都没有提及转染后是用转染的这同一批细胞同时做pcr,wb,cck8,凋亡,细胞周期;还是说这次转染只做pcr或wb,再转染一次做cck8或细胞周期。剩下的功能试验均同前,转染一次做一次?我养的是悬浮细胞,转染后做cck8这些功能试验前需要离心换液吗?跪谢解答!
悬浮细胞的转染方法:(以下内容转自生物帮资讯)
DXY721认为:
悬浮细胞和贴壁细胞在转染过程中差别不大,主要差别在于转染后的筛选,当然如果你做的是瞬时转染就不存在筛选的问题了。
其实转染的过程很简单,问题是能不能转的进去的,转染率能有多少,转进去是否可以稳定表达目的蛋白等等。
我们也是用脂质体做悬浮细胞的转染,说明书上都有具体的操作过程,将脂质体和目的基因按比例混合,然后加到细胞悬液里就OK了,说的简单,实际上还是有一些细节要注意的,比如脂质体和目的基因混合的比例,转染的细胞数,细胞的代数,细胞的状态,有的还要求在转染的前一天传代一次,不过不要怕,这些在脂质体说明书上都有明确的说明,按照说明书做就可以了。
jinghuanlv认为:
悬浮细胞和贴壁细胞转染还是有很大不同的。
脂质体转染的原理基于电荷吸引原理,先形成脂质体-DNA复合物,散布在细胞周围,然后通过细胞的内吞作用,将目的基因导入细胞内,而脂质体复合物与贴壁细胞的接触机会比悬浮细胞高出很多倍,所以,脂质体转染时悬浮细胞的转染效率要明显低于贴壁细胞。
我们实验室转染悬浮细胞是用的电穿孔法,目前为止,悬浮细胞转染的最好方法还是电转,我们实验室用的电转仪是Bio-Rad的,使用条件是电压250V,电容975uF,效果不错,不妨一用。

