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: C₂₂H₂₉N₇O₅•₂HCl

For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.

* Refer to Certificate of Analysis for lot specific data (including water content).

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Puromycin dihydrochloride: sc-108071. Cultured HeLa cells treated with 0µg/ml (A)...

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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.

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 :

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

Download SDS (MSDS)

Certificate of Analysis

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Puromycin dihydrochloride (CAS 58-58-2) Product Citations

See how others have used Puromycin dihydrochloride (CAS 58-58-2). Click on the entry to view the PubMed entry .

Citations 1 to 10 of 318 total

PMID: # 35586315 Elías-Villalobos, A.|Duncan, C.|Mata, J.|Helmlinger, D.| et al. 2022. STAR Protoc. 3: 101373.

Citations 1 to 10 of 318 total

Hello, first of all thank you for your answer.I have many problems with my cell line. I used different concentrations of puromycin from 0,15 ug/uL to 12 ug/ml and I always have a few cells in the dish. Should I still increase the concentration? Thank you

Asked by: frfra

Thank you for your question. Please contact technical service directly for troubleshooting advice on your experiment.

Answered by: Tech Support Europe

Date published: 2021-03-10

Hello, I want to use DMSO to dissolve puromycin but I would like to know if I have to filter it or not. Thank you!

Asked by: frfra

Thank you for your question. This product is not sterile, so filtration is recommended for use in cell culture.

Answered by: Tech Support Europe

Date published: 2021-02-24

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.