Puromycin
| Product | Unit size | Cat. code | Docs. | Qty. | Price | |
|---|---|---|---|---|---|---|
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Puromycin (solution) Selective antibiotic for the Pac gene |
Show product |
100 mg (10 x 1 ml) 500 mg (50 x 1 ml) 500 mg (1 x 50 ml) |
ant-pr-1
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Selection antibiotic: endotoxin tested, sterile reagent
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.
Reference:
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.
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Specifications
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 :
Contents
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.
Details
WORKING CONCENTRATIONS
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. Rössger 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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FAQs
What is the best way to resuspend Puromycin?
We recommend to resuspend Puromycin in HEPES buffer (5 mM, pH 7.3)
We have noticed a crystalline precipitate of Puromycin when thawing a vial left at -20°C, is this normal?
Yes, during storage a crystalline precipitate of Puromycin may form. If this occurs, heat the product at 37°C for 30 minutes until the crystalline precipitate disappears. The formation of a crystalline precipitate does not affect the activity of the product.
What is the preferred storage method for Puromycin?
Puromycin is shipped at room temperature. Upon receipt, it should be stored at 4°C for short term storage or -20°C for long term storage. Avoid repeated freeze-thaw cycles. The expiry date is specified on the product label. Once opened it remains stable for 1 year at -20°C providing you avoid repeated freeze-thaw cycles, and it can be kept at 4°C for 3 months provided it is kept in sterile conditions.
What is the selection concentration of Puromycin in mammalian cells?
The working concentration of Puromycin in mammalian cell lines varies between 1-10 μg/ml. In an initial experiment we recommend to determine the optimal concentration of puromycin required to kill your host cell line. Please note that Puromycin quickly kills eukaryotic cells that do not contain the pac gene.
For more detailed information, please refer to the technical data sheet for this product.
What is the selection concentration of Puromycin in E.coli?
Puromycin has low activity in E. coli. However, puromycin-resistant transformants can be selected in LB agar medium, with the pH adjusted to 7.5-8, supplemented with 100-125 μg/ml of Puromycin.
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