Puromycin dihydrochloride CAS: 58-58-2
MF: C22H29N7O5•2HCl
MW: 544.43

Puromycin dihydrochloride (CAS 58-58-2)

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Alternate Names: Puromycin dihydrochloride is also known as Stylomycin hydrochloride
Application: Puromycin dihydrochloride is an antibiotic used for selection and maintenance of cell lines expressing a transfected pac gene
CAS Number: 58-58-2
Purity: ≥98%
Molecular Weight: 544.43
Molecular Formula: C22H29N7O52HCl
仅供科研使用。不可用于诊断或治疗。
* 参考分析证明 大量特定数据 (包括水 含量).
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Puromycin dihydrochloride, also known as Stylomycin hydrochloride, is an aminonuclease antibiotic used for selection and maintenance of cell lines expressing a transfected pac gene, whose product, puromycin-N-acetyl-transferase, inactivates puromycin via acetylation.
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.


参考文献

1. Brunetti, M., et al. 1995. Blood. 86: 4199-4205. PMID: 7492778
2. Constam, D.B., et al. 1995. J. Biol. Chem. 270: 26931-26939. PMID: 7592939
3. Sugita, M., et al. 1995. Zool. Sci. 12: 419-425. PMID: 8528014
4. Tabuchi, I. 2003. Biochem. Biophys. Res. Commun. 305: 1-5. PMID: 12732187

Usage :
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.
Appearance :
Powder
Physical State :
Solid
Derived From :
Streptomyces alboniger
Solubility :
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).
Storage :
Desiccate at -20° C
Melting Point :
178-180° C
仅供科研使用。不可用于诊断或治疗。
WGK Germany :
3
RTECS :
AU7355000
PubChem CID :
64806
Merck Index :
14: 7943
MDL Number :
MFCD00012691
EC Number :
200-387-8
Beilstein Registry :
3853613
SMILES :
CN(C)C1=NC=NC2=C1N=CN2[[email protected]]3[[email protected]@H]([[email protected]@H]([[email protected]](O3)CO)NC(=O)[[email protected]@H](CC4=CC=C(C=C4)OC)N)O.Cl.Cl

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Citations 1 to 10 of 229 total

PMID: # 31138784  Starr, CR. et al. 2019. Cell Death Dis. 10: 409.

PMID: # 30587510  McCaughey, J. et al. 2019. J. Cell Biol. 218: 929-948.

PMID: # 31278301  Koedoot, E.|Fokkelman, M.|Rogkoti, VM.|Smid, M.|van de Sandt, I.|de Bont, H.|Pont, C.|Klip, JE.|Wink, S.|Timmermans, MA.|Wiemer, EAC.|Stoilov, P.|Foekens, JA.|Le Dévédec, SE.|Martens, JWM.|van de Water, B.| et al. 2019. Nat Commun. 10: 2983.

PMID: # 31269628  Hutchinson, SA.|Lianto, P.|Moore, JB.|Hughes, TA.|Thorne, JL.| et al. 2019. Int J Mol Sci. 20:

PMID: # 31569690  2019. Cells. 8:

PMID: # 31545550  2019. FEBS J.

PMID: # 30838870  Wu, M.|Feng, Y.|Ye, GX.|Han, YC.|Wang, SS.|Ni, HF.|Wang, FM.|Gao, M.|Lv, LL.|Liu, BC.| et al. 2019. Am. J. Physiol. Renal Physiol. 316: F1006-F1015.

PMID: # 30579117  Wen, J.|Bai, H.|Chen, N.|Zhang, W.|Zhu, X.|Li, P.|Gong, J.| et al. 2019. Mol. Immunol. 106: 53-62.

PMID: # 31123703  Di Rienzo, M.|Antonioli, M.|Fusco, C.|Liu, Y.|Mari, M.|Orhon, I.|Refolo, G.|Germani, F.|Corazzari, M.|Romagnoli, A.|Ciccosanti, F.|Mandriani, B.|Pellico, MT.|De La Torre, R.|Ding, H.|Dentice, M.|Neri, M.|Ferlini, A.|Reggiori, F.|Kulesz-Martin, M.|Piacentini, M.|Merla, G.|Fimia, GM.| et al. 2019. Sci Adv. 5: eaau8857.

PMID: # 30700707  LaMonte, GM.|Orjuela-Sanchez, P.|Calla, J.|Wang, LT.|Li, S.|Swann, J.|Cowell, AN.|Zou, BY.|Abdel-Haleem Mohamed, AM.|Villa Galarce, ZH.|Moreno, M.|Tong Rios, C.|Vinetz, JM.|Lewis, N.|Winzeler, EA.| et al. 2019. Nat Commun. 10: 488.

Citations 1 to 10 of 229 total
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