
Description
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Transfection Reagent for bEnd.3 Cell Line (Brain Endothelioma Cells, CRL-2299)
- A nanoparticle-based liposome formulation
- Transfection protocols provided for transfection of proteins, DNA, mRNA, siRNA, shRNA and microRNA
- Transfection Enhancer reagent provided with the kit
- Produce higher level of recombinant protein expression with minimal disruption of normal cell function
- Generate physiologically relevant data you can trust
- Effective for plasmid DNA/siRNA co-transfection
- Easy-to-use transfection protocol with reproducible results
- Low cytotoxicity
- Download in vitro bEnd3 transfection protocol: [PDF]
- Download bEnd-3 CRISPR/Cas9 transfection protocol: [PDF]
- Download PowerPoint presentation for bEnd3 cells transfection kit: [PPT]
- Developed and manufactured by Altogen Biosystems
Transfection Efficiency:
Reagent exhibits at least 70% transfection efficiency of siRNA delivery. Transfection efficiency was determined by qRT-PCR.
Transfection Protocol and MSDS:
Download Altogen Biosystems Transfection Protocol: [PDF]
Download MSDS: [PDF]
bEnd.3 Cell Line:
Stroke is a life-threatening disease that affects arteries in the brain and can be caused by the blockage of the arteries by a blood clot, resulting in seizures. bEnd.3 is a brain endothelial cell line taken from a mouse’s (Mus musculus) brain/cerebral cortex. As endothelial cells, they make up the interior surface of blood vessels and lymphatic vessels, which circulate blood. These cells form the structure that not only helps carry nutrients to the brain but also removes waste. The bEnd.3 cell line is used in a variety of molecular and cell biology research. Study of this cell could be useful for medical research related to the flow of blood, providing a test subject for ways to treat clots and congestion, which can cause seizures and other serious health effects. Altogen Biosystems offers an advanced liposome formulation bEnd-3 transfection reagent kit that provides high transfection efficiency with bEnd-3 cells and is a helpful tool for biomedical research applications related to the flow of blood and blood flow obstruction.
Data:
Figure 1. GAPD mRNA levels were quantified using real-time RT-PCR in the bEnd3 cells transfected with siRNAs targeting GAPD or non-silencing siRNA. Forty-eight hours post-transfection, the cells were harvested and analyzed by real-time RT-PCR for GAPD mRNA expression levels. Data were normalized against the 18S rRNA signal. Control samples were either mock-transfected or untreated. Values are normalized to untreated sample. Data are means ± SD (n=5).
Figure 2. Protein expression of GAPDH in bEnd-3 cells. DNA plasmid expressing GAPDH or siRNA targeting GAPDH were transfected into bEnd-3 cells following Altogen Biosystems transfection protocol. At 72 hours post-transfection the cells were analyzed by Western Blot for protein expression levels (normalized by total protein, 10 µg of total protein loaded per each well). Untreated cells used as a negative control.
Selected in vivo transfection product citations (ALTOGEN® IN VIVO Transfection Kits used in the following publications):
- Nature Biotechnology. 2011 29(4):341-5. Delivery of siRNA to the mouse brain by … Alvarez-Erviti et al [PDF]
- Jounal of Biological Chemistry. 2012 287(4):2907. Chaperoning of mutant p53 protein … Gogna et al [PDF]
- PLoS Pathogens. 2012 8(8) Uridine composition of the poly-U/UC tract of HCV RNA … Schnell et al [PDF]
- J Proteome Res. 2012(11) Retinal proteome analysis in a mouse model of oxygen-induced … Kim et al [PDF]
- J Transl Med. 2010 15;8:133. Prevention of hyperglycemia-induced myocardial apoptosis … Zhang et al [PDF]
bEnd-3 Transfection Reagent
Altogen Biosystems:
Altogen Biosystems is a life sciences company that manufactures over 100 cell type specific and preoptimized transfection kits, electroporation buffers, and targeted in vivo delivery kits and reagents. Advanced formulation of reagents and optimized transfection protocols provide highly efficient intracellular delivery of biomolecules (proteins, DNA, mRNA, shRNA and siRNA, and small molecule compounds). Read more about transfection technology at Altogen’s Transfection Resource.
Altogen Research Services:
Altogen Labs provides GLP-compliant contract research studies for pre-clinical research, IND applications, and drug development. Biology CRO services include: Xenograft models (30+), development of stable cell lines, ELISA assay development, cell-based and tissue targeted RNAi studies, safety pharm/tox assays, and other studies (visit AltogenLabs.com).
Volume Options:
- 0.5 ml (Catalog #6145)
- 1.5 ml (Catalog #6146)
- 1.5 ml CRISPR (Catalog #2113)
- 8.0 ml (Catalog #6147)
AltogenBiosystems是一家开发和制造用于生命科学研究,药物发现和开发的转染试剂盒的生物技术公司。转染试剂盒针对特定癌细胞系和原代细胞培养进行了优化,可将生物分子有效递送到靶组织中。通过先进的试剂配方和优化的转染方案实现体外(癌细胞系)和体内(动物组织靶向试剂、癌细胞系)递送货物分子,包括质粒DNA,各种类型的RNA(mRNA,siRNA,shRNA,microRNA),蛋白质和小分子研究。
Altogen生命科学公司致力于研发,生产和销售特定细胞系的转染试剂,用于细胞间生物分子的传递,并通过对转染试剂类型的设计将siRNA和质粒DNA有效地转入不同的细胞系和原代细胞内。Altogen公司开发的聚合物,脂质体,纳米粒子为基础的转染技术分别针对分子生物学,组合化学,和细胞生物学而分别应用。Altogen定制服务提供符合GLP要求定制研究服务,包括代稳定的细胞系,细胞银行和冷冻保存,焦磷酸测序,克隆,RNA干扰(RNAi)和基因沉默服务,发展分析,siRNA文库筛选,并转染服务。稳定的肿瘤细胞株和原代细胞的产生,可以是非常昂贵和费时。该公司的细胞培养科学家的细胞株的选择,无论是利息或shRNA表达载体的稳定表达的基因改造。标准的RNAi技术服务,包括设计与合成的siRNA的利益,验证siRNA的沉默效率,siRNA转染条件的优化,使高效的基因沉默细胞系或原代培养细胞的靶基因。转染培养细胞的瞬时或稳定的引入外源性分子和遗传物质(即RNA或DNA),通常是在生物实验室用来研究基因功能,基因表达的调节,生化映射,突变分析,和蛋白质的生产。科学家利用各种载体分子,这种分子,使质粒DNA(PDNA),信使RNA(mRNA),短干扰RNA(siRNA),小分子RNA(miRNA)的,并进入肿瘤细胞株和原代细胞的蛋白质的基因交付。不幸的是,无单提货的方法或转染试剂,可以适用于所有类型的细胞,细胞的细胞毒性和转染效率显着不同,取决于试剂,协议,并正在利用细胞类型。Altogen生物系统公司提供超过60种类型的细胞的预优化转染试剂盒。纳米粒子,脂质和聚合物基ALTOGEN®在体内转染试剂,使交付功能的RNA和DNA分子在体内。PEG脂质体在体内输送系统减少由于PEG修饰的先天免疫反应,并提供高效的siRNA转染的DNA,并在体内的蛋白质。由科学“杂志(2010年12月17日):PEG脂质体在体内转染试剂盒siRNA的特色Altogen生物系统功能的特定细胞系转染试剂盒
120+细胞转染试剂和活体组织靶向试剂盒制造商AltogenBiosystems是一家生物技术公司,开发和制造用于生命科学研究、药物发现和开发的转染试剂盒。Altogen®体内转染试剂可有效地将生物分子导入靶组织。细胞转染试剂盒针对特定的癌细胞系和原代细胞进行了优化。通过先进的试剂配方和优化的转染方案实现货物分子(DNA、RNA、蛋白质)的高效传递。AltogenBiosystems利用高分子化学、分子和细胞生物学的专业知识,开发了新的体内外给药技术。转染是将外源分子导入培养细胞中,常用于研究基因功能、基因表达调控、生化定位和蛋白质生产。不幸的是,由于细胞毒性和转染效率的差异很大,并且取决于所使用的试剂、方案和细胞类型,因此没有一种单一的传递方法或转染试剂可应用于所有类型的细胞。AltogenBiosystems为120多个癌细胞系和原代细胞类型提供优化的转染试剂盒和电穿孔产品。体内转染试剂可实现组织靶向给药。Altogen的转染试剂盒包括用于体外(癌细胞系)和体内(用于动物研究的组织靶向试剂)转染的转染增强剂试剂和转染复合物冷凝器。Altogen实验室提供符合GLP的实验室合同研究服务。我们的生物CRO服务包括异种移植物的疗效、IND应用的pharm/tox研究和安全性测试、分析开发(ELISA、IC-50、qPCR)、90多个异种移植物动物模型、RNAi和基因沉默服务。Altogen的细胞培养科学家通过在28天内培育出稳定的细胞系,将选择的细胞系转化为稳定表达感兴趣的基因。
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有战友对比过罗氏X-tremeGENEDNATransfectionReagent和lipofectamine3000的优劣吗?求解答
而且直接使用自制的PEI非常便宜,在293上远比商品化的脂质体要好。
另外,如果你们实验室确实钱多,不怕花钱,建议你取用Promega的FugenHD,那个转染效率比脂质体更好,而且毒性小,至于价格。。。。。。。。。。。也更高。。。。。。。。。
另外,你说漂浮的细胞有表到GFP,那个不一定是真的GFP,很多时候
内毒素是革兰氏阴性菌细胞壁(cellwall)上的特有成分,主要是脂多糖中的类脂A,在细菌被裂解时被释放出来,由于其化学结构和特性,在质粒的纯化过程中很容易混入质粒DNA一同提取出来。内毒素的存在会严重的影响质粒转染细胞的效率,此外会激活造血细胞(如B细胞、巨噬细胞等)的非特异免疫反应,造成实验的假阳性,所以转染级质粒的提取纯化必须去除内毒素。
—DNA转染试剂Polyjet-适用于普遍的哺乳动物细胞
LipoD293-适用于悬浮细胞的转染(包括昆虫细胞SF9)-对于普遍的哺乳动物细胞具有更优异的转染效率
—DNAandsiRNA转染试剂
Lipojet-适用于大多数哺乳动物细胞的转染-低毒性(无需换液)-用量少-DNA/RNA共转染(co-transfection)-效果优于Lipofectamine2000
—siRNA转染试剂
GenMute-适用于大多数哺乳动物细胞转染-低毒性(无需换液)-用量少-DNA/siRNA共转染(co-transfection)PepMute-适用于普遍的哺乳动物细胞的转染
如题,PolyplusTransfection转染试剂在中国区的代理商有哪些?求推荐1-2个靠谱的,谢谢!
想用RNAiMAX或者lipo2000转染siRNA,但是看了下转染试剂的说明书和锐博的siRNA说明书,觉得分别对siRNA的用量描述差别挺大的呀,不知道到底该参考哪个呢。
以24孔板为例在siRNA说明书中写到,siRNA终浓度是50nM的话,加入浓度为20μM的siRNA1.25ul,每孔体积是500ul,那这样的话,每孔最终siRNA的量是25pmol。
但是在RNAiMAX或者是lipo2000说明书中,一个写的每孔siRNA用量是5pmol,一个是500ng,这与siRNA厂家所提供的量相差也太多了吧。
到底该看哪一个呢。
ps.一旦siRNA的量和体积确定下来之后,转染试剂的量和siRNA1:1的加就可以了吗?
请各位大神解答。
1.siRNA说明书中的用量,红线圈出
2.RNAIMAX说明书中siRNA的用量。
3.lipo2000说明书中siRNA用量

