Puromycin dihydrochlorideAn aminonuclease antibiotic used for selection and maintenance of cell lines expressing a transfected pac gene
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Puromycin dihydrochloride (CAS 58-58-2)

Puromycin dihydrochloride | CAS 58-58-2 is rated 4.9 out of 5 by 14.
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Synonym: Stylomycin hydrochloride
Application: 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: C22H29N7O5•2HCl
* Refer to Certificate of Analysis for lot specific data (including water content).
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Puromycin 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.
This product has been cell culture tested and is recommended for selection of stably transfected cells following transfection with Santa Cruz Biotechnology shRNA or Lentiviral particles.
BACKGROUND: Puromycin 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. Toxic to both eukaryotic and prokaryotic cells. Believed to act as an acyl-tRNA analogue causing premature chain termination, it has been shown to arrest cells in G2/M phase.


References

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
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.
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[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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Certificate of Analysis

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Puromycin dihydrochloride  Product Citations

See how others have used Puromycin dihydrochloride. Click on the entry to view the PubMed entry .

Citations 1 to 10 of 196 total

PMID: # 30910993  Fulzele, S.|Mendhe, B.|Khayrullin, A.|Johnson, M.|Kaiser, H.|Liu, Y.|Isales, CM.|Hamrick, MW.| et al. 2019. Aging (Albany NY). 11: 1791-1803.

PMID: # 30578966  Giudetti, AM.|De Domenico, S.|Ragusa, A.|Lunetti, P.|Gaballo, A.|Franck, J.|Simeone, P.|Nicolardi, G.|De Nuccio, F.|Santino, A.|Capobianco, L.|Lanuti, P.|Fournier, I.|Salzet, M.|Maffia, M.|Vergara, D.| et al. 2019. Biochim Biophys Acta Mol Cell Biol Lipids. 1864: 344-357.

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

PMID: # 30443895  Uhl, J.|Gujarathi, S.|Waheed, AA.|Gordon, A.|Freed, EO.|Gousset, K.| et al. 2019. J Cell Commun Signal. 13: 209-224.

PMID: # 31114073  He, L.|Zhou, J.|Chen, M.|Lin, CS.|Kim, SG.|Zhou, Y.|Xiang, L.|Xie, M.|Bai, H.|Yao, H.|Shi, C.|Coelho, PG.|Bromage, TG.|Hu, B.|Tovar, N.|Witek, L.|Wu, J.|Chen, K.|Gu, W.|Zheng, J.|Sheu, TJ.|Zhong, J.|Wen, J.|Niu, Y.|Cheng, B.|Gong, Q.|Owens, DM.|Stanislauskas, M.|Pei, J.|Chotkowski, G.|Wang, S.|Yang, G.|Zegarelli, DJ.|Shi, X.|Finkel, M.|Zhang, W.|Li, J.|Cheng, J.|Tarnow, DP.|Zhou, X.|Wang, Z.|Jiang, X.|Romanov, A.|Rowe, DW.|Wang, S.|Ye, L.|Ling, J.|Mao, J.| et al. 2019. Nat Mater. 18: 627-637.

PMID: # 30881116  Chiang, KC.|Huang, ST.|Wu, RC.|Huang, SC.|Yeh, TS.|Chen, MH.|Hsu, JT.|Chen, LW.|Kuo, SF.|Chueh, HY.|Juang, HH.|Hung, SI.|Yeh, CN.|Pang, JS.| et al. 2019. Cancer Manag Res. 11: 1893-1905.

PMID: # 31545550  2019. FEBS J.

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

Citations 1 to 10 of 196 total

Hi, following reconstitution in water as a 50 mg/ml stock, can it be stored at 4°C for regular short-term use? If so, how long is it stable at 4°C? Should it be added fresh to media, or can it be added directly to media stored at 4°C (as in pen/strep)?

Asked by: AlineS
We always recommend using solutions the same day they are made, but if that is not possible, the solution should be stable for up to 3 months stored at -20ºC.
Answered by: Technical Support
Date published: 2019-11-06

Was there any animal component in contact, while preparing the Puromycin dihydrochloride (catalog number - CAS 58-58-2)

Asked by: CG_Pharma
Thank you for your question. This product is synthetic and was not exposed or derived from any animal components.
Answered by: Technical Support
Date published: 2019-07-25

What concentration of the puromycin is the good initially start for testing CRISPR?Cas9 KO plasmid?

Asked by: Rose H
The working puromycin concentration for mammalian cell lines ranges from 1–10 µg/ml. Prior to using the puromycin antibiotic (sc-108071), titrate the selection agent to determine the optimal concentration for target cell line. Use the lowest concentration that kills 100% of non-transfected cells in 3–5 days from the start of puromycin selection.
Answered by: Technical Support
Date published: 2019-02-10

Hello. I would like to know how long the puromycin powder stored at -20ºC remains stable. Beforehand thank you very much.

Asked by: Pablo_Morgado
Stable for at least 1 year after receipt when stored, as supplied, at -20°C. Stock solutions are stable for up to 3 months at -20°C.
Answered by: Technical Support
Date published: 2019-02-10

what is the purity of PUROMYCIN DIHYDROCHLORIDE ？

Asked by: Kenny123
Thank you for your question. Each lot of this chemical comes with a lot-specific Certificate Of Analysis which typically describes the percent purity as well as the method used to determine the purity. You may request a lot-specific Certificate Of Analysis on our website. Alternatively, please contact our Technical Service Department to inquire about the purity of our most recent lot. Please call our Asian Support office toll-free at 800-820-8626, email asia@scbio.cn, or contact us by live chat.
Answered by: Technical Support
Date published: 2018-11-22

How to make 5 ug/mL solution in water?

Asked by: Jz28sail
Puromycin dihydrochloride is soluble in water (50 mg/ml). To make a solution of 5 micrograms of Puromycin dihydrochloride in water, we recommend you dissolve the specified quantity of Puromycin dihydrochloride in filtered water. The solution should appear clear, colorless to slightly yellow, and free from particulates. We recommend filtering the solution with a 0.22µm filter.
Answered by: Chemical Support 5
Date published: 2018-11-18

Can this product be stored in solution?

Asked by: hawkeye11
Thank you for your question. This chemical, Puromycin dihydrochloride (CAS 58-58-2), can be stored in solution at -20ºC, for up to 3 months.
Answered by: Chemical Support 1
Date published: 2018-06-21

Is this product cell culture grade?

Asked by: hawkeye11
Thank you for your question. This chemical, Puromycin dihydrochloride (CAS 58-58-2), is not considered cell culture grade.
Answered by: Chemical Support 1
Date published: 2019-05-20
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Rated 5 out of 5 by from Puromycin Great quality antibiotic, we used it for selection of the transducer suspension cells. Worked well.
Date published: 2018-03-28
Rated 4 out of 5 by from Great Until now, it works very well. I recommend it for you
Date published: 2018-03-20
Rated 5 out of 5 by from puromycin Puromycin used for cell line selection. it is very good
Date published: 2016-12-19
Rated 5 out of 5 by from Helps a lot The effect in my experiment is good using this product, fast screening results are stable, and easy to identify. It helps a lot in improving my experiment plans.
Date published: 2016-12-06
Rated 5 out of 5 by from GOOD FUCTION I used this for the cell screening and have a good fuction.
Date published: 2016-11-25
Rated 5 out of 5 by from You make me believe Used several times and it performs great. No complains
Date published: 2016-11-04
Rated 5 out of 5 by from Tested a small amount initially for CRISPR Tested a small amount initially for CRISPR experiments, got great results, purchased bulk amounts.
Date published: 2015-10-19
Rated 5 out of 5 by from We used it to select for KO in our cell line We used it to select for KO in our cell line and it worked really well.
Date published: 2015-10-12
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