Polysciences PEI溶液配制说明书

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Polysciences PEI溶液配制说明书

——线性聚乙烯亚胺, #23966,1mg/ml转染试剂的配制

转染试剂线性PEI溶液的配制

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，Ethyl ether 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。

参考文献：

1.聚乙烯亚胺（PEI）、线性 (1 mg/mL). Cold Spring Harbor Protocols 2008, pdb.rec11323–pdb.rec11323 (2008). doi:10.1101/pdb.rec11323.

 

 

 

 

Catalog# 23966 –Polyethylenimine, Linear, MW 25000 (PEI 25K)

Transfection Reagent Preparation and Storage Recommendations

These guidelines are our recommendations based off research, testing, and customer feedback. There are alternative methods available, but we suggest following these recommendations for easy adoption and consistently high performance. These methods are adapted from Cold Spring Harbor Protocols [1], an excellent secondary resource for transient transfection literature.

Reagent Preparation (1 mg/mL)

Materials:

1. 1g PEI 25K (Polysciences Catalog# 23966).

2. 1L Milli-Q® water, water for injection (WFI), or comparable biological-grade water

3. 12 M Hydrochloric Acid (HCl)

4. 10 M Sodium Hydrochloride (NaOH)

5. Disposable 0.1-0.2µm PES vacuum sterile-filter.

6. Sterile HDPE or polypropylene storage vials for reagent aliquots.

Equipment

1. 1L glass beaker

2. 1L glass graduated cylinder

3. Calibrated pH meter

4. 2 x disposable 1mL plastic pipettes

5. Stir plate

6. PTFE coated stir bar

7. Vacuum pump

Method:

1. Dispense 1g of PEI 25K into beaker and suspend in 900 mL of water.

2. Add stir bar and set stirring to produce a small vortex.

3. Add hydrochloric acid dropwise until pH is < 2.0.

4. Cover top of beaker and stir for up to 3 hours until powder fully dissolves.

5. Add sodium hydroxide dropwise until pH is 6.9-7.1.

6. Transfer solution to graduated cylinder and add water until the total volume is 1L.

7. Sterile-filter through vacuum membrane.

8. Aliquot as desired and store at -20°C.

Reagent Storage

Frozen aliquots can be stored at -20°C for up to one year. Aliquots can be thawed and kept at 4°C for up

to two weeks, but should never be re-frozen.

Powder Storage

Unopened powder has a shelf life of two years from date of receipt. It can be kept at room temperature

until use.

References

1. Polyethylenimine (PEI), linear (1 mg/mL). Cold Spring Harbor Protocols 2008, pdb.rec11323–pdb.rec11323 (2008). doi:10.1101/pdb.rec11323

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