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polysciences公司23966中英文說明
美國polysciences公司成立于1961年,主要生產(chǎn)和經(jīng)營LIFE SCIENCES生命科學(xué)(膠體金,不含甲醛的甲醇等),病理學(xué)和顯鏡學(xué) 微球和粒子和磁珠各種單體和聚體等產(chǎn)品。polysciences公司是世界上zui大zui全的多聚化合物供應(yīng)商,同時(shí)是世界*的免疫學(xué),組織化學(xué)試劑耗材供應(yīng)商.
轉(zhuǎn)染試劑線性PEI溶液的配制(1mg/ml)
聚乙烯亞胺(Polyethylenimine,PEI)是聚乙烯亞胺是一種具有較高的陽離子電荷密度的有機(jī)大分子,每相隔二個(gè)碳個(gè)原子,即每“第三個(gè)原子都是質(zhì)子化的氨基氮原子,使得聚合物網(wǎng)絡(luò)在任何pH下都能充當(dāng)有效的“質(zhì)子海綿”(proton sponge)體。PEI,可用作轉(zhuǎn)染試劑,它可以縮聚DNA分子形成顆粒并轉(zhuǎn)染真核細(xì)胞。有實(shí)驗(yàn)證明,分子量為25000的線性PEI即Polysciences 23966轉(zhuǎn)染效率zui高。
下面將主要介紹polysciences 線性PEI(#23966)的產(chǎn)品特性及其溶液的配制過程。
1. 產(chǎn)品特性:
產(chǎn)品名稱:線性聚乙烯亞胺(PEI)
CAS號(hào) | 9002-98-6,26913-06-4 | 分子式 | (CH2CH2NH)n | 分子量 | 25000 |
熔點(diǎn) | 73-75°C | 外觀 | 白色或黃色固體 | 結(jié)構(gòu)式 |
|
溶解性 | 溶于熱水,低pH值的冷水,甲醇和乙醇;不溶于:苯,aether和丙酮 | 儲(chǔ)存 | 室溫 |
2. 溶液配制
此指導(dǎo)說明書是經(jīng)polysciences公司研究,測(cè)試及其客戶反饋的后編寫的。下面推薦一種簡(jiǎn)單適用而的方法,講述如何配制濃度為1mg/mL的轉(zhuǎn)染試劑的配制。
材料:
1g #23966、1L 純水(Milli-Q®純水,注射用水(WFI),或類似的生物級(jí)水)、12 M鹽酸、10 M氫氧化鈉、一次性0.1-0.2µm PES真空無菌過濾器、無菌高聚乙烯或聚丙烯試劑瓶。
器材:
1 L 玻璃燒杯、1 L玻璃量筒、核準(zhǔn)pH計(jì)、2支1 mL塑料移液管、托盤、聚四氟乙烯(PTFE)攪拌棒、真空泵
配制過程:
1g #23966——900mL水溶解——攪拌、加熱(60-80°C)——加HCl調(diào)pH到2.0左右——蓋上燒杯,攪拌3h(直至粉末*溶解)——冷卻至室溫——加NaOH調(diào)pH到6.9-7.1——移液到量筒,加水補(bǔ)足至1L——真空過濾消毒——分裝,-20°C保存。
備注:加熱或加酸都有助于PEI溶解。當(dāng)然酸可能不用加那么多,詳細(xì)用量可試加一點(diǎn)攪拌看看,足夠溶解就行,不一定需要加到pH那么低。
試劑儲(chǔ)存:
冷凍部分溶液可在-20°C保存長達(dá)一年;部分解凍的溶液可在4°C保存2周,不可反復(fù)凍融。
粉末儲(chǔ)存:
未開封的粉末有效期兩年。可在室溫下保存直至使用。
polysciences公司23966中英文說明
英文說明:
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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