Polyethylenimine Linear, MW 25000 (PEI 25K)

線性化聚乙烯亞胺PEI 25000

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搜索關鍵詞：

PEI25K;PEI 25000;PEI 40K;線性化聚乙烯亞胺PEI；PEI轉染試劑；PEI 25000；CAS：9002-98-6, 26913-06-4；Polysciences;

產品信息

【溫馨提示】：見我司整理的線性化聚乙烯亞胺PEI 25000/ PEI40000轉染試劑專題。

產品描述

線性化聚乙烯亞胺PEI 25，000，一種高電荷陽離子聚合物，非常容易結合于高電荷陰離子基質。工業應用上，線性化PEI可改善負電荷染料的物理外觀，以調整染料的化學特性和增強染料與固相基質的黏附能力，常用作黏附增強劑，作用同多聚賴氨酸（poly-lysine）；科研應用上，線性化PEI能與DNA或其他負電荷生物大分子簡單結合，正是這一特性，使得成為一種非常行之有效的載體運輸介質（Vector carrier），即轉染試劑。

線性化聚乙烯亞胺PEI 25000（Polyethylenimine Linear, MW 25000）是一款優秀、低成本、值得信賴的瞬時性轉染試劑。在HEK293和CHO表達系統中，PEI在寬廣的生產規模內（從96孔板到100L生物反應器）能提供連續性的高基因表達。

產品特性

1）CAS NO.：9002-98-6, 26913-06-4

2）分子量：25,000

3）外觀：白色至黃色固體

4）溶解性：溶于熱水、冷水（低pH），甲醇和乙醇；不溶于苯、二乙醚、丙酮

5）化學結構式：

保存與運輸方法

保存：室溫保存，或可置于4℃延長保存周期，2年有效。

運輸：室溫運輸。

儲存液配制

1）于1L玻璃燒杯，將1g PEI25K粉末加入900ml Milli-Q®超純水或其他相當級別的生物用水中。

2）將磁性轉子放入燒杯內，打開攪拌模式使產生小型漩渦。邊攪拌邊逐滴加入鹽酸（12M）調節pH，直至pH＜2.0。

3）蓋住燒杯口，持續攪拌直至粉末完全溶解（溶解時間高達3h）。整個過程pH＜2.0?！咀⒁狻浚嚎赡軙嬖谝恍┬±w維狀顆粒不能溶解，這是正?，F象。

4）邊攪拌邊逐滴加入NaOH（10M）調節pH，直至到6.9-7.1。

5）將溶液轉入量筒內，加入水定容到1L，用一次性0.1-0.2μm PES真空過濾器過濾除菌，即得到1mg/ml的儲存液。

6）根據單次用量將儲存液分裝，保存在-20℃，高達1年穩定。儲存液再次融化后，可置于4℃保存，高達2周穩定，但絕不可重新凍存。

注意事項

1）對于本品的轉染方法，可參考我司提供的PEI 25K Transfection Reagent線性化聚乙烯亞胺轉染試劑

（貨號：MX2204-1ML），或參考以下幾篇文獻：

①Durocher, 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).

② 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.

③ 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).

④ 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).

2）為了您的安全和健康，請穿實驗服并戴一次性手套操作。

PEI25K客戶使用案例（逐步增加中）

圖片描述：以人結腸癌細胞（HCT116）為對象，按照質粒DNA：PEI 2,5000（MX2202-250MG）=1:4的比例制備轉染混合物并轉染進入HCT116細胞，36h后于熒光顯微鏡下觀察轉染效率（GFP，綠色熒光蛋白）。

【數據來源：廈門大學 于老師】

相關產品

 — —Written/Edited by V. Shallan【版權歸MKBio懋康所有】

上海懋康生物科技有限公司是一家涉足于生命科學和生物技術領域研究的試劑、儀器和實驗室消耗品與實驗服務工作，主要從事細胞生物學、植物學、分子生物學、免疫學、生物化學、蛋白組學。生物制藥與診斷試劑研發生產等領域。 本公司秉承“以人為本，以誠為信、合同守信”的經營理念。堅持"品質保障"的原則為廣大客戶提供優質產品。

引用文獻：

[1] Liu Bin et al. AAV-Containing Exosomes as a Novel Vector for Improved Gene Delivery to Lung Cancer Cells. Front. Cell Dev. Biol., 13 August 2021

AAV and AAVExo Production and Purification

AAVs were produced by double transfection of HEK 293T cells as described previously (Rapti et al., 2012). Briefly, cells were cultured in a T175 flask with culture medium. When 60–70% confluency was achieved, cell culture medium was replaced with transfection reagent, which was made by mixing 50 μg of the helper plasmid, 17 μg of the transgene plasmid, and 233 μl of polyethylenimine (1 mg/ml, linear, MW ～25,000; Cat. No. MX2202;

Maokang, Shanghai, China) in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 2% fetal bovine serum (FBS) and streptomycin–penicillin. The cells were collected 3 days later at 300 g for 10 min (with cell-free supernatant saved for AAVExo purification) and resuspended in 10 ml of lysis buffer (150 mmol/l sodium chloride, 50 mmol/l Tris–HCl, pH = 8.5), subjected to three freeze–thaw cycles and treated with 1,500 U of benzonase nuclease (Cat. No. MP1509-25KU; Maokang, China) in the presence of 1 mmol/l magnesium chloride for 1 h at 37°C. Cellular debris was removed by centrifugation for 10 min at 5,000 g (Avanti J-E with a JA-25.50 rotor, Beckman Coulter, Brea, CA, United States). The virus was purified by a four-step iodixanol gradient centrifugation [5.8 ml of 15%, 3.9 ml of 25%, 3.1 ml of 40%, and 3.1 ml of 60% iodixanol (Optiprep, Cat. No. D1556; Sigma-Aldrich), overlayed with 10 ml of cell lysate in lysis buffer] in a 70Ti rotor (Beckman Coulter, Brea, CA) at 68,000 rpm for 1 h using polycarbonate bottles (Cat. No. 355618; Beckman Coulter). The 40–60% interphase of the gradient was collected, and the buffer was exchanged using a Vivaspin20 column with 100,000 MWCO (Sartorius, G?ttingen, Germany) in sterile phosphate-buffered saline (PBS).

[2] Zhang X, Hong K, Sun Q, Zhu Y, Du J. Bioreducible, arginine-rich polydisulfide-based siRNA nanocomplexes with excellent tumor penetration for efficient gene silencing. Biomater Sci. 2021 Jul 27;9(15):5275-5292. doi: 10.1039/d1bm00643f. PMID: 34180478.

PEI 25K was purchased from Maokang Biological Technology Co

[3] Zuo Z, Li L, Yan X, Zhang L. Glucose Starvation Causes ptauS409 Increase in N2a Cells Through ATF3/PKAcα Signaling Pathway. Neurochem Res. 2022 Nov;47(11):3298-3308. doi: 10.1007/s11064-022-03686-x. Epub 2022 Jul 20. PMID: 35857208.

Cell transfection was performed using PEI 25,000 (MKbio, Shanghai) for plasmid DNA transfection (DNA: PEI = 1:5).