Puromycin dihydrochloride (Synonyms: CL13900) |
| Catalog No.GC16384 |
Puromycin dihydrochloride is produced by Streptomyces alboniger, a grampositive actinomycete, through a series of enzymatic reactions.
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 58-58-2
Sample solution is provided at 25 µL, 10mM.
Puromycin dihydrochloride is produced by Streptomyces alboniger, a grampositive actinomycete, through a series of enzymatic reactions.[1] Puromycin dihydrochloride included a nucleoside covalently bound to an amino acid, mimicking the 30 end of aminoacylated tRNAs that participate in delivery of amino acids to elongating ribosomes.[2] It inhibits the growth of animal cells and blocks protein synthesis by binding to 80S ribosomes at low doses.[3]
In vitro study determined the optimal concentration of Puromycin dihydrochloride for selecting EGFPac-transfected cells by performing a Puromycin dihydrochloride resistance test. The puromycin-resistant gene (termed pac) encoding puromycin N-acetyl transferase was isolated from Streptomyces aboniger. If pac is introduced and expressed in animal cells, the cells can survive in the presence of Puromycin dihydrochloride. Results ahowed that it could successfully produce a somatically cloned transgenic piglet using recombinant cells obtained after gene transfer of a transgene (carrying both EGFP and pac expression units) and subsequent in vitro selection with a low concentration (2 mg/ml) of puromycin.[3]
In vivo study was conducted to determine the surface sensing of translation (SUnSET) technique could be used to measure the protein synthesis in whole tissues. Since there is currently an intense interest in identifying the molecular mechanisms that regulate skeletal muscle protein synthesis. It allows for the visualization and quantification of protein synthesis and eliminates the need for generating radioactive tissues/animals. This study also determined that the surface sensing of translation could detect relatively acute changes in protein synthesis in the absence of changes in rRNA as well as detect not only increases but also decreases in protein synthesis in vivo. [4]
References:
[1]. Tercero JA, Espinosa JC, Lacalle RA, Jiménez A. The biosynthetic pathway of the aminonucleoside antibiotic puromycin, as deduced from the molecular analysis of the pur cluster of Streptomyces alboniger. J Biol Chem 1996;271(3):1579–90.
[2]. Aviner R. et al. The science of puromycin: From studies of ribosome function to applications in biotechnology. Comput Struct Biotechnol J. 2020 Apr 24;18:1074-1083.
[3]. Watanabe S, Iwamoto M, et al. A novel method for the production of transgenic cloned pigs: electroporation-mediated gene transfer to non-cultured cells and subsequent selection with puromycin. Biol Reprod. 2005 Feb;72(2):309-15.
[4]. Goodman CA, Mabrey DM, et al. Novel insights into the regulation of skeletal muscle protein synthesis as revealed by a new nonradioactive in vivo technique. FASEB J. 2011 Mar;25(3):1028-39.
| Cell experiment [1]: | |
|
Cell lines |
Fetal porcine somatic cells |
|
Preparation Method |
Cells were seeded in 24-well plates at a density of 2.5 x 104 cells per well and cultured in medium containing 0.5–6 μg/ml puromycin dihydrochloride. Stock solution (10 mg/ml) of puromycin dihydrochloride was prepared by dissolving puromycin dihydrochloride in distilled water at the appropriate concentration. Media containing variable amounts of puromycin dihydrochloride were freshly prepared by adding the appropriate volume of puromycin dihydrochloride stock solution. |
|
Reaction Conditions |
0.5–6 μg/ml for 7 d |
|
Applications |
Puromycin dihydrochloride is an antibiotic that inhibits growth of animal cells and blocks protein synthesis by binding to 80S ribosomes at low doses. To determine the optimal concentration of puromycin dihydrochloride for selecting EGFPac-transfected cells, a puromycin dihydrochloride resistance test was performed with fetal porcine somatic cells. The puromycin-resistant gene (termed pac) encoding puromycin N-acetyl transferase was isolated from Streptomyces aboniger. If pac is introduced and expressed in animal cells, the cells can survive in the presence of puromycin dihydrochloride. |
| Animal experiment [2]: | |
|
Animal models |
Female FVB/N mice, 8–10 weeks old. |
|
Preparation Method |
Puromycin dihydrochloride was dissolved in 100 μl of PBS. Mice were housed under a 12-h light/dark cycle with ad libitum access to food and water unless otherwise stated. Mice were anesthetized with an intraperitoneal injection of ketamine (100 mg/kg) and xylazine (10 mg/kg) before all surgical procedures. |
|
Dosage form |
Puromycin dihydrochloride was intraperitoneal injected to mice with a concentration of 0.040 μmol/g. |
|
Applications |
The antibiotic puromycin dihydrochloride (a structural analog of tyrosyl-tRNA), and anti-puromycin antibodies could be used to detect the amount of puromycin incorporation into nascent peptide chains as well as to measure changes in protein synthesis in cell cultures. |
|
References: [1]. Watanabe S, Iwamoto M, et al. A novel method for the production of transgenic cloned pigs: electroporation-mediated gene transfer to non-cultured cells and subsequent selection with puromycin. Biol Reprod. 2005 Feb;72(2):309-15. [2]. Goodman CA, Mabrey DM, et al. Novel insights into the regulation of skeletal muscle protein synthesis as revealed by a new nonradioactive in vivo technique. FASEB J. 2011 Mar;25(3):1028-39. |
|
| Cas No. | 58-58-2 | SDF | |
| Synonyms | CL13900 | ||
| 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 | ||
| Canonical SMILES | O[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 | ||
| Formula | C22H29N7O5.2HCl | M.Wt | 544.43 |
| Solubility | ≥ 27.2mg/mL in DMSO, ≥ 99.4mg/mL in Water | Storage | Store at -20°C |
| General tips | Please select the appropriate solvent to prepare the stock solution according to the
solubility of the product in different solvents; once the solution is prepared, please store it in
separate packages to avoid product failure caused by repeated freezing and thawing.Storage method
and period of the stock solution: When stored at -80°C, please use it within 6 months; when stored
at -20°C, please use it within 1 month. To increase solubility, heat the tube to 37°C and then oscillate in an ultrasonic bath for some time. |
||
| Shipping Condition | Evaluation sample solution: shipped with blue ice. All other sizes available: with RT, or with Blue Ice upon request. | ||
| Prepare stock solution | |||
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1 mg | 5 mg | 10 mg |
| 1 mM | 1.8368 mL | 9.1839 mL | 18.3678 mL |
| 5 mM | 0.3674 mL | 1.8368 mL | 3.6736 mL |
| 10 mM | 0.1837 mL | 0.9184 mL | 1.8368 mL |
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- Purity: >98.00%
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