Puromycin dihydrochloride [58-58-2]
Référence sc-108071
Conditionnement : 25mg
Marque : Santa Cruz Biotechnology
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Puromycin dihydrochloride (CAS 58-58-2)
ACCÈS RAPIDE AUX LIENS
Puromycin dihydrochloride has been cell culture tested and is recommended for selection of stably transfected cells following transfection with Santa Cruz Biotechnology shRNA or Lentiviral particles. Puromycin dihydrochloride is an antibiotic substance produced by the soil actinomycete Streptomyces alboniger which induces apoptosis in cells by interfering with RNA function, leading to inhibition of protein synthesis. Puromycin dihydrochloride is thought to act as an acyl-tRNA analogue causing premature chain termination. Toxic to both eukaryotic and prokaryotic cells, Puromycin dihydrochloride has also been shown to arrest cells in G2/M phase.
References:
- Next-generation protein-handling method: puromycin analogue technology. | Tabuchi, I. 2003. Biochem Biophys Res Commun. 305: 1-5. PMID: 12732187
- Immune surveillance for ERAAP dysfunction. | Nagarajan, NA. and Shastri, N. 2013. Mol Immunol. 55: 120-2. PMID: 23433779
- Pathophysiology and treatment of focal segmental glomerulosclerosis: the role of animal models. | de Mik, SM., et al. 2013. BMC Nephrol. 14: 74. PMID: 23547922
- The hypusine-containing translation factor eIF5A. | Dever, TE., et al. 2014. Crit Rev Biochem Mol Biol. 49: 413-25. PMID: 25029904
- Genetic associations and functional characterization of M1 aminopeptidases and immune-mediated diseases. | Agrawal, N. and Brown, MA. 2014. Genes Immun. 15: 521-7. PMID: 25142031
- Therapeutic target for nephrotic syndrome: Identification of novel slit diaphragm associated molecules. | Fukusumi, Y., et al. 2014. World J Nephrol. 3: 77-84. PMID: 25332898
- Regulatory T cells and minimal change nephropathy: in the midst of a complex network. | Bertelli, R., et al. 2016. Clin Exp Immunol. 183: 166-74. PMID: 26147676
- Antibiotics and RNase P. | Drainas, D. 2016. Antibiotics (Basel). 5: PMID: 27164152
- Chloramphenicol Derivatives as Antibacterial and Anticancer Agents: Historic Problems and Current Solutions. | Dinos, GP., et al. 2016. Antibiotics (Basel). 5: PMID: 27271676
- Combined use of electron microscopy and intravital imaging captures morphological and functional features of podocyte detachment. | Burford, JL., et al. 2017. Pflugers Arch. 469: 965-974. PMID: 28664407
- Transient and stable vector transfection: Pitfalls, off-target effects, artifacts. | Stepanenko, AA. and Heng, HH. 2017. Mutat Res Rev Mutat Res. 773: 91-103. PMID: 28927539
- Spontaneous and glucocorticoid-induced apoptosis in human mature T lymphocytes. | Brunetti, M., et al. 1995. Blood. 86: 4199-205. PMID: 7492778
- Puromycin-sensitive aminopeptidase. Sequence analysis, expression, and functional characterization. | Constam, DB., et al. 1995. J Biol Chem. 270: 26931-9. PMID: 7592939
- Puromycin induces apoptosis of developing chick sympathetic neurons in a similar manner to NGF-deprivation. | Sugita, M., et al. 1995. Zoolog Sci. 12: 419-25. PMID: 8528014
Utilisation :
For the selection of stably transfected cells following transfection with Santa Cruz Biotechnology shRNA or Lentiviral particles, proceed with puromycin selection as follows:
NOTE: The working puromycin concentration for selection in mammalian cell lines ranges from 1-10 µg/ml. Prior to using the puromycin antibiotic, titrate the selection agent to determine the optimal concentration for target cell line (see below). Use the lowest concentration that kills 100% of non-transfected cells in 3-5 days from the start of puromycin selection.
•48 hours post-transfection, aspirate the medium and replace with fresh medium containing puromycin at the appropriate concentration.
•Approximately every 2-3 days, aspirate and replace with freshly prepared selective media.
•Monitor the cells daily. Puromycin selection requires a minimum of 48 hours. Optimum effectiveness should be reached within 3-10 days.
•Assay transfected cells.
NOTE: Lot-to-lot variations in potency exist for all selection antibiotics, each new lot of puromycin should be titrated.
Titrating puromycin for selecting transfected cell lines:
•Plate 2 x 105 cells in each well of a 6-well plate containing 3 ml of the appropriate complete medium plus increasing concentrations of puromycin (i.e., 0, 1.0, 2.5, 5.0, 7.5, and 10.0 µg/ml)
•Replace with fresh selective medium after 2 days to remove dead cells.
•Examine the wells for viable cells every two days.
•Monitor the cells daily and observe the percentage of surviving cells. Optimum effectiveness should be reached in 1-4 days.
•The minimum antibiotic concentration to use is the lowest concentration that kills 100% of the cells in 3-5 days from the start of puromycin selection.
Apparence :
Powder
État Physique :
Solid
Dérivé de :
Streptomyces alboniger
Solubilité :
Soluble in water (50 mg/ml), ethanol (~1 mg/ml), DMSO (~13 mg/ml), DMF (~14 mg/ml), and PBS, pH 7.2 (~10 mg/ml).
STOCKAGE :
Desiccate at -20° C
Point de fusion :
178-180° C
WGK Allemagne :
3
RTECS :
AU7355000
PubChem CID :
64806Index Merck :
14: 7943
Numéro MDL :
MFCD00012691
Numéro CE :
200-387-8
Registre Beilstein :
3853613
SMILES :
CN(C)C1=NC=NC2=C1N=CN2[C@H]3[C@@H]([C@@H]([C@H](O3)CO)NC(=O)[C@@H](CC4=CC=C(C=C4)OC)N)O.Cl.Cl
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SAMPLE Certificate of Analysis (COA)
Puromycin dihydrochloride | CAS 58-58-2 SAMPLE Certificate of AnalysisCERTIFICAT D'ANALYSE
Numéro de catalogue
Numéro de Lot
SAMPLE Certificate of Analysis (COA)
| Test | Specification | Results |
|---|---|---|
| Appearance | White to off-white powder | |
| Identification | Positive | Complies |
| Purity (HPLC) | ≥ 98.0% | 99.1% |
| Purity (TLC) | NP, CH2Cl2, methanol 7:3RP C18, methanol, H2O 9:1 | > 99%> 99% |
| Melting Point | 168 - 182°C | 176 - 180°C |
| Solubility | Clear and colorless or light yellow solution (50mg/mL in H2O) | Clear colorless solution |
| Water Content | ≤ 12.0% | 6.2% |
| Country of Origin | Israel |
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