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polysciences公司23966中英文说明
美国polysciences公司成立于1961年,主要生产和经营LIFE SCIENCES生命科学(胶体金,不含甲醛的甲醇等),病理学和显镜学 微球和粒子和磁珠各种单体和聚体等产品。polysciences公司是世界上zui大zui全的多聚化合物供应商,同时是世界*的免疫学,组织化学试剂耗材供应商.
转染试剂线性PEI溶液的配制(1mg/ml)
聚乙烯亚胺(Polyethylenimine,PEI)是聚乙烯亚胺是一种具有较高的阳离子电荷密度的有机大分子,每相隔二个碳个原子,即每“第三个原子都是质子化的氨基氮原子,使得聚合物网络在任何pH下都能充当有效的“质子海绵”(proton sponge)体。PEI,可用作转染试剂,它可以缩聚DNA分子形成颗粒并转染真核细胞。有实验证明,分子量为25000的线性PEI即Polysciences 23966转染效率zui高。
下面将主要介绍polysciences 线性PEI(#23966)的产品特性及其溶液的配制过程。
1. 产品特性:
产品名称:线性聚乙烯亚胺(PEI)
CAS号 | 9002-98-6,26913-06-4 | 分子式 | (CH2CH2NH)n | 分子量 | 25000 |
熔点 | 73-75°C | 外观 | 白色或黄色固体 | 结构式 |
|
溶解性 | 溶于热水,低pH值的冷水,甲醇和乙醇;不溶于:苯,aether和丙酮 | 储存 | 室温 |
2. 溶液配制
此指导说明书是经polysciences公司研究,测试及其客户反馈的后编写的。下面推荐一种简单适用而的方法,讲述如何配制浓度为1mg/mL的转染试剂的配制。
材料:
1g #23966、1L 纯水(Milli-Q®纯水,注射用水(WFI),或类似的生物级水)、12 M盐酸、10 M氢氧化钠、一次性0.1-0.2µm PES真空无菌过滤器、无菌高聚乙烯或聚丙烯试剂瓶。
器材:
1 L 玻璃烧杯、1 L玻璃量筒、核准pH计、2支1 mL塑料移液管、托盘、聚四氟乙烯(PTFE)搅拌棒、真空泵
配制过程:
1g #23966——900mL水溶解——搅拌、加热(60-80°C)——加HCl调pH到2.0左右——盖上烧杯,搅拌3h(直至粉末*溶解)——冷却至室温——加NaOH调pH到6.9-7.1——移液到量筒,加水补足至1L——真空过滤消毒——分装,-20°C保存。
备注:加热或加酸都有助于PEI溶解。当然酸可能不用加那么多,详细用量可试加一点搅拌看看,足够溶解就行,不一定需要加到pH那么低。
试剂储存:
冷冻部分溶液可在-20°C保存长达一年;部分解冻的溶液可在4°C保存2周,不可反复冻融。
粉末储存:
未开封的粉末有效期两年。可在室温下保存直至使用。
polysciences公司23966中英文说明
英文说明:
PEI 25K is a powerful, trusted, and cost-effective transient transfection reagent. In HEK293 and CHO expression systems, PEI offers consistently high gene expression on a wide scale (96 well plates up to 100 L bioreactors). Each year, more researchers and companies turn to polysciences PEI to gain a critical edge in their work. Relative to most other options, using PEI to prepare transfection reagents in-house can offer as much as a 40% reduction in total transfection costs.
CAS#:
9002-98-6, 26913-06-4
Formula:
(CH2CH2NH)n
Molecular Weight:
25,000
Melting Point:
73-75º
Soluble In:
hot water, cold water at low pH, methanol, and ethanol.
Insoluble In: benzene, ethyl ether and acetone
Appearance:
White to yellow solid
Reference(s):
Siegman S, Truong NF, Segura T. (2015). Encapsulation of PEGylated low-molecular-weight PEI polyplexes in hyaluronic acid hydrogels reduces aggregation. Acta Biomater. in Press, doi:10.1016/j.actbio.2015.09.020
Baranyi, L. et al. Rapid Generation of Stable Cell Lines Expressing High Levels of Erythropoietin, Factor VIII, and an Antihuman CD20 Antibody Using Lentiviral Vectors. Human Gene Therapy Methods 24, 214–227 (2013). doi:10.1089/hgtb.2013.002
Wulhfard, S., Baldi, L., Hacker, D. L. & Wurm, F. Valproic acid enhances recombinant mRNA and protein levels in transiently transfected Chinese hamster ovary cells. Journal of Biotechnology 148, 128–132 (2010). doi:10.1016/j.jbiotec.2010.05.003
Thomas M, Lu JJ, Ge Q, Zhang C, Chen J, Klibanov AM. (2005). Full deacylation of polyethylenimine dramatically boosts its gene delivery efficiency and specificity to mouse lung. Proc Natl Acad Sci U S A.102(16):5679-84.
DuRocher, Y., Perret, S. & Kamen, A. High-level and high-throughput recombinant protein production by transient transfection of sUSPension-growing human 293-EBNA1 cells. Nucleic acids research 30, E9 (2002). doi:10.1093/nar/30.2.e9
TSCA
Hazards:
Unknown
Handling:
Gloves & fume hood
Storage:
Store at room temperature
MSDSMaterial Safety Datasheet 23966 (PDF)
DATATransfection Reagent Preparation and Storage Recommendations
DATAExample COA
LITBSE/TSE Statement
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