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  6. Puromycin, Sterile

Puromycin, Sterile 

Cat. No.: HY-K1057
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Culture Assay

Puromycin is an aminonucleoside antibiotic produced by Streptomyces alboniger. It inhibits protein synthesis by disrupting peptide transfer on ribosomes, causing premature chain termination during translation. It can kill most gram-positive bacteria and various animal or insect cells.

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65 Publications Citing Use of MCE Puromycin, Sterile

Cell Imaging/Staining
IF
Cell Proliferation/Viability Assay
WB

    Puromycin, Sterile purchased from MCE. Usage Cited in: Adv Mater. 2025 Dec 1:e16597.  [Abstract]

    The stable 4T1-Vector and 4T1-P16INK4a cell lines were established via lentiviral transduction with Vector and pP16INK4a plasmids, respectively, followed by selection with 5 µg/mL Puromycin. Representative images of SA-β-gal staining and FCM analysis of 4T1-Vector and 4T1-P16 INK4a cells.

    Puromycin, Sterile purchased from MCE. Usage Cited in: Cell Mol Immunol. 2025 Aug 29.  [Abstract]

    For transduction, intestinal organoids at a low passage number and 2d post-passaging were incubated with pretreated lentivirus (LRIG1−/− or control) in culture medium containing polybrene. The suspension was centrifuged at 37 °C and 1000 ×g for 30 min and incubated at 37 °C for 4 h. After transduction, the organoids were cultured in medium supplemented with 1 μg/mL Puromycin for selection. Representative images of intestinal organoids cultured with supernatant.

    Puromycin, Sterile purchased from MCE. Usage Cited in: Cell Mol Immunol. 2025 Aug 29.  [Abstract]

    Lenti-CRISPR viruses were generated via triple-plasmid transfection. Briefly, murine LRIG1 sgRNAs were designed with the Synthego CRISPR design tool. The oligos were synthesized, annealed, and cloned and inserted into the LentiCRISPR-LRIG1 vector, followed by Sanger sequencing verification. HEK293T cells were cotransfected with LentiCRISPR-LRIG1 or LentiCRISPR-Control along with the lentiviral second-generation helper plasmids psPAX2 and pCMV-VSV-G via PEI transfection reagent. The culture supernatants harvested at 48 h and 72 h posttransfection were filtered, concentrated with PEG8000 and finally frozen at −80 °C. For transduction, intestinal organoids at a low passage number and 2d $ost-passaging were incubated with pretreated lentivirus (LRIG1−/− or control) in culture medium containing polybrene. The suspension was centrifuged at 37 °C and 1000 ×g for 30 min and incubated at 37 °C for 4 h. After transduction, the organoids were cultured in medium supplemented with 1 μg/mL Puromycin for selection.

    Puromycin, Sterile purchased from MCE. Usage Cited in: Nat Commun. 2024 Dec 5;15(1):10599.  [Abstract]

    Cells were incubated with Puromycin (10 µg/mL) at 37 °C for 30 min, permeabilized and fixed for intracellular staining with AF488-conjugated anti-puromycin (1:100). Puromycin intake levels were detected by flow cytometry.

    Puromycin, Sterile purchased from MCE. Usage Cited in: Nat Commun. 2025 Nov 26;16(1):10514.  [Abstract]

    For the stable expression of mCherry-PCBP2, SH-SY5Y and SH-SY5Y-APP cells were transduced with LV6-mCherry-PCBP2 and LV6-mCherry vectors. Forty-eight hours post-transduction, the cells were transferred to 10 cm dishes and supplemented with 2 μg/mL Puromycin. Clones expressing mCherry-PCBP2 were identified and selected using fluorescence microscopy.

    Puromycin, Sterile purchased from MCE. Usage Cited in: Adv Sci (Weinh). 2025 Aug 13:e17432.  [Abstract]

    To induce K‐TM silence in Jurkat T cells, single‐guide RNA was designed and inserted into pLX‐hPGK‐EGFP vector and constructed into lentivirus using the lentiviral packaging method described above. Transfection of sgRNA was performed with polyjet transfection reagent and cells were selected with Puromycin for 5 days and then used for cytotoxicity assay. As expected, Jurkat‐K‐TMKO T cells elicited stronger cytotoxicity against tumor cells compared to the control.

    • Description

    • Storage

    • Protocol

    • Components

    • Documentation

    Description
    & Advantages

    Puromycin is an aminonucleoside antibiotic produced by Streptomyces alboniger. It inhibits protein synthesis by disrupting peptide transfer on ribosomes, causing premature chain termination during translation. It can kill most gram-positive bacteria and various animal or insect cells.

    Resistance to Puromycin is conferred by the puromycin N-acetyl-transferase gene (pac) from Streptomyces alboniger. Therefore, Puromycin is a popular and effective antibiotic for selecting vectors bearing the pac gene in mammalian cell lines and E. coli.

    Storage

    -20°C; 2 years.
    Protocol

    1. The working concentration of Puromycin varies with cell type, media, growth conditions and cell metabolic rate.

    2. Before stable transfected cell lines can be selected, the optimal Puromycin concentration needs to be determined by performing a kill curve titration.

    1) Seed the parental cell line in a suitable culture plate at a cell density of 20-25% and incubate the cells for 24 hours at 37°C.

    2) Remove medium and then add medium with various concentrations of Puromycin and incubate at 37°C.

    3) Refresh the selective medium every 2-3 days and observe the percentage of surviving cells over time.

    4) Determine the lowest concentration of antibiotic that kills a large majority of the cells within 7-10 days. This concentration should be used for selection of a stable transfected cell line.
    Components
    Components HY-K1057-1 mL HY-K1057-5 mL
    Puromycin, Sterile (10 mg/mL) 1 mL 5 mL
    Documentation

    SDS (168 KB)
    Help & FAQs
    • Do most proteins show cross-species activity?

      Species cross-reactivity must be investigated individually for each product. Many human cytokines will produce a nice response in mouse cell lines, and many mouse proteins will show activity on human cells. Other proteins may have a lower specific activity when used in the opposite species.

    Keywords:

    Cell Culture AntibioticsSelection AntibioticsAntibioticCell contaminationbacterial contaminationPuromycin Dihydrochloride

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