更新時間：2022-01-28
Puromycin is an aminonucleoside antibiotic produced by Streptomyces alboniger. It specifically inhibits peptidyl transfer on both prokaryotic and eukaryotic ribosomes. This antibiotic inhibits the ……
Puromycin is an aminonucleoside antibiotic produced by Streptomyces alboniger. It specifically inhibits peptidyl transfer on both prokaryotic and eukaryotic ribosomes. This antibiotic inhibits the growth of Gram positive bacteria and various animal and insect cells.
Puromycin can also be used in some particular conditions for the selection of E. coli transformants. Resistance to puromycin is conferred by the Pac gene encoding a puromycin N-acetyl-transferase [1].
Animal cells are generally sensitive to concentrations from 1 to 10 µg/ml.
1. Lacalle R. et al., 1989. Molecular analysis of the pac gene encoding a puromycin N-acetyl transferase from Streptomyces alboniger. Gene. 79:375-80.
Product concentration: 10 mg/ml
CAS number: 58-58-2
Quality Control: Each lot is thoroughly tested to ensure the absence of lot-to-lot variation.
Purity: ≥ 98% (HPLC)
Endotoxin level: < 5 EU/mg
Physicochemical characterization: pH, appearance
Cell-culture tested: potency validated in puromycin-sensitive and puromycin?resistant mammalian cell lines
Non-cytotoxicity of trace contaminants: absence of long-term effects confirmed in puromycin-resistant cells
Formula: C22H29N7O5 • 2HCl
Molecular weight: 544.3
Structure :
Puromycin dihydrochloride is supplied as a sterile filtered solution at 10 mg/ml in HEPES buffer.
This product is available in three pack sizes:
ant-pr-1: 10 x 1 ml (100 mg)
ant-pr-5: 50 x 1 ml (500 mg)
ant-pr-5b: 1 x 50 ml (500 mg)
Puromycin is shipped at room temperature.
Upon receipt it should be stored at 4°C or -20°C.
Puromycin is a harmful compound. Refer to safety data sheet for handling instructions.
The working concentrations of puromycin for mammalian cell lines range from 1 to 10 μg/ml.
In a starting experiment we recommend to determine optimal concentrations of antibiotic required to kill your host cell line.
Puromycin quickly kills eukaryotic cells that do not contain the pac gene. Dying cells detach from the plates, allowing easy and early identification of transformant clones.
Suggested working conditions for selection in some mammalian cells are listed below.
Cell line | Medium | Puromycin conc | References |
|---|---|---|---|
| B16 (Mouse melanocytes) | RPMI | 1-2 μg/ml | 1, 2 |
| HEK293 (Human embryonic kidney cells) | DMEM | 0.5-10 μg/ml | 3-5 |
| HeLa (Human uterine cells) | DMEM | 1-10 μg/ml | 5, 6 |
| MEF (Mouse fibroblasts) | DMEM | 1-5 μg/ml | 5 |
References:
1. Furge KA. et al., 2001. Suppression of Ras-mediated tumorigenicity and metastasis through inhibition of the Met receptor tyrosine kinase. PNAS 98:10722-7.
2. Díaz J. et al., 2014.Rab5 is required in metastatic cancer cells for Caveolin-1-enhanced Rac1 activation, migration and invasion.. J Cell Sci. 127:2401-6.
3. R?ssger K. et al., 2013. Reward-based hypertension control by a synthetic brain-dopamine interface. PNAS, 110:18150-5.
4. Schmitter D. et al., 2006. Effects of Dicer and Argonaute down-regulation on mRNA levels in human HEK293 cells. Nucleic Acids Res. 34:4801-15.
5. Kamer I. et al., 2005. Proapoptotic BID Is an ATM effector in the DNA-damage response Cell. 122:593-603.
6. Charnaux N. et al., 2005. RANTES (CCL5) induces a CCR5-dependent accelerated shedding of syndecan-1 (CD138) and syndecan-4 from HeLa cells and forms complexes with the shed ectodomains of these proteoglycans as well as with those of CD44. Glycobiology. 15:119-30.
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