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Puromycin dihydrochloride

现货
Catalog No.
B7587
氨酰tRNA类似物,用于表达pac基因的稳定细胞株的筛选
组合的产品项目
规格价格库存 数量
10mM (in 1mL H2O)
¥ 600.00
现货
20mg
¥ 700.00
现货

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Background

Puromycin dihydrochloride [1] is an analog of aminoacyl-tRNA. Puromycin dihydrochloride allows selection for cells expressing the resistance gene puromycin N-acetyl-transferase (PAC) and is useful in CRISPR/Cas9 mammalian gene editing by selecting for successful Cas9-induced knock-in with puromycin resistance gene. It inhibits the incorporation of aminoacyl-tRNA into the C-terminal on a synthesizing polypeptide, resulting in the premature termination of the polypeptide chain.

Puromycin is toxic to the growth of various eukaryote cells including mammalian cells. Concentrations of puromycin sufficient to inhibit the cell growth of mammalian cells range from 0.5-10 μg/ml. IC90s for puromycin to inhibit the growth of Plasmodium falciparum and Giardia lamblia are 60 ng/ml and 54 μg/ml, respectively [2]. Aminoacyl-tRNA is used as the material for elongation of a synthesizing polypeptide on a ribosome [2].

When treated with puromycin dihydrochloride at different concentrations, the growth rates of T. thermophila changed. In the first 24 h, puromycin dihydrochloride at a concentration of 50 µg/ml reduced the growth rate by 80%, but did not completely block the cell growth; until 72 h, there was a gradual cell number increase. At 100 μg/ml, puromycin dihydrochloride completely blocked the cell growth; in the first 48 h under this condition, almost all of the cells died, surviving cells grew rapidly after 48 h. Puromycin dihydrochloride at 150 μg/ml completely inhibited the cell growth for 72 h. By 72 h, the majority of cells died, and then surviving cells grew. Puromycin dihydrochloride at 200 μg/ml made almost all the cells die by 48 h, and hence no survivors appeared [2].

In animals of 25 days old, 180 or 120 min of previous exposure to puromycin dihydrochloride inhibited subsequent amino acid transport. In animals of 50 days old, however, puromycin dihydrochloride failed to inhibit α-aminoisobutyric acid uptake [3].

References:
[1].  Mi. Sundaralingami and S. K. Arora. Stereochemistry of nucleic acids and their constituents. IX. The conformation of the antibiotic puromycin dihydrochloride pentahydrate. Proc. N. A. S., 1969, 64(3):1021-6.
[2].  Masaaki Iwamoto, ChieMori, Yasushi Hiraoka, et al. Puromycin resistance gene as an effective selection marker for ciliate Tetrahymena. Gene, 2014, 534:249–255.
[3].  Elsas L.J. II, MacDonell R.C. Jr. and Rosenberg L.E. Influence of age on insulin stimulation of amino acid uptake in rat diaphragm. J. Biol. Chem., 1971, 246(21):6452-6459.

Chemical Properties

Physical AppearanceA solid
StorageStore at -20°C
M.Wt544.43
Cas No.58-58-2
FormulaC22H29N7O5.2HCl
Solubility≥27.2mg/mL in DMSO, ≥99.4mg/mL in H2O
Chemical Name(S)-2-amino-N-((2S,3R,4S,5R)-5-(6-(dimethylamino)-9H-purin-9-yl)-4-hydroxy-2-(hydroxymethyl)tetrahydrofuran-3-yl)-3-(4-methoxyphenyl)propanamide dihydrochloride
SDFDownload SDF
Canonical SMILESO[C@H]1[C@H]([C@@H](CO)O[C@H]1N2C3=NC=NC(N(C)C)=C3N=C2)NC([C@H](CC(C=C4)=CC=C4OC)N)=O.Cl.Cl
运输条件试用装:蓝冰运输。 其他可选规格:常温运输或根据您的要求用蓝冰运输。
一般建议为了使其更好的溶解,请用37℃加热试管并在超声波水浴中震动片刻。不同厂家不同批次产品溶解度各有差异,仅做参考。若实验所需浓度过大至产品溶解极限,请添加助溶剂助溶或自行调整浓度。

试验操作

细胞实验 [1]:

细胞系

嗜热四膜虫

制备方法

在DMSO中的溶解度大于10 mM。若配制更高浓度的溶液,一般步骤如下:请将试管置于37 °C加热10分钟和/或将其置于超声波浴中震荡一段时间。原液于-20 °C可放置数月。

反应条件

0 ~ 200 μg/mL;0 ~ 72小时

实验结果

200 μg/mL Puromycin Dihydrochloride于48小时后杀死所有嗜热四膜虫。此后,没有再出现嗜热四膜虫。

动物实验 [2]:

动物模型

25天龄和50天龄的小鼠

给药剂量

0.2 mg/g;腹腔注射

实验结果

Puromycin Dihydrochloride是一种自噬诱导物,能提高游离核糖体的含量。给予Puromycin Dihydrochloride,于30 ~ 60分钟后,高达70%的核糖体恢复其游离形式。

其它注意事项

请于室内测试所有化合物的溶解度。虽然化合物的实际溶解度可能与其理论值略有不同,但仍处于实验系统误差的允许范围内。

References:

[1]. Masaaki Iwamoto, ChieMori, Yasushi Hiraoka, et al. Puromycin resistance gene as an effective selection marker for ciliate Tetrahymena. Gene, 2014, 534:249–255.

[2]. Réz G, Kiss A, Bucsek MJ, Kovács J. Attachment of ribosomes to endoplasmic membranes in mouse pancreas. Degranulation in vivo caused by the inducers of autophagocytosis neutral red, vinblastine, puromycin, and cadmium ions, and prevention by cycloheximide. Chem Biol Interact. 1976 Apr;13(1):77-87.

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