包装 | 价格(元) |
10mM (in 1mL DMSO) | 电议 |
20mg | 电议 |
50mg | 电议 |
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 mg/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 | Cells were incubated with puromycin for 7 days. Prepared Puromycin dihydrochloride was stored at 48 ℃. The concentration of puromycin dihydrochloride should be lower than 2 μg/ml. |
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 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. |
文献引用 | |
产品描述 | 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: |