Puromycin, Sterile
Based on 73 publication(s) in Google Scholar
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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Storage :
-20°C; 2 years.
- SDS (168KB)
- COA
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.
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.
Publications
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Journal Impact Factor
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Most Recent
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Adv MaterSenescence Reprogramming Unleashes Tumor Immune Surveillance via Coordinated Gene Modulation. [Abstract]2025 Dec 1:e16597. PMID: 41324273
Puromycin, Sterile purchased from MedChemExpress. 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.
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Cell Mol ImmunolMacrophage-derived VISTA engages with LRIG1 and hinders gut epithelial repair in colitis. [Abstract]2025 Aug 29. PMID: 40883589
Puromycin, Sterile purchased from MedChemExpress. 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 MedChemExpress. 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.
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Nat Commun2025 Nov 26;16(1):10514. PMID: 41298370
Puromycin, Sterile purchased from MedChemExpress. 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.
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Nat CommunAberrant METTL1-mediated tRNA m7G modification alters B-cell responses in systemic autoimmunity in humans and mice. [Abstract]2024 Dec 5;15(1):10599. PMID: 39638793
Puromycin, Sterile purchased from MedChemExpress. 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.
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Cell Discov2024 Nov 5;10(1):112. PMID: 39500876
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Adv Sci (Weinh)HERV-K TM Subunit Elicits CD8+ T Cell Anergy and Tumor Immune Evasion via Targeting CD3 Coreceptor ε in AML and PDAC. [Abstract]2025 Aug 13:e17432. PMID: 40801282
Puromycin, Sterile purchased from MedChemExpress. 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.
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Cell Rep MedA combined "eat me/don't eat me" strategy based on exosome for acute liver injury treatment. [Abstract]2025 Apr 15;6(4):102033. PMID: 40120577
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Cell Death Dis2025 Apr 10;16(1):271. PMID: 40210616
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J Transl MedChemically synthesized osteocalcin alleviates NAFLD via the AMPK-FOXO1/BCL6-CD36 pathway. [Abstract]2024 Aug 22;22(1):782. PMID: 39175012
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OncogeneKPNA2 promotes osteosarcoma progression by regulating the alternative splicing of DDX3X mediated by YBX1. [Abstract]2025 Jul;44(26):2186-2200. PMID: 40216969
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ApoptosisUSP9X regulates copper-induced CASP9/CASP3/GSDME-dependent pyroptosis by deubiquitinating JAK1. [Abstract]2026 Jun 18;31(7):177. PMID: 42315669
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Int J Biol MacromolBAG6 promotes hepatocellular carcinoma growth via enhancing PLK1-mediated aerobic glycolysis. [Abstract]2025 Dec 11;338(Pt 1):149653. PMID: 41390024
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Cancer Biol TherRET fusion partners dictate oncogenic potential in undifferentiated spindle cell sarcomas. [Abstract]2026 Dec 31;27(1):2683190. PMID: 42240026
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Int J Mol SciRegulation of PHD2 by HIF-1α in Erythroid Cells: Insights into Erythropoiesis Under Hypoxia. [Abstract]2025 Jan 17;26(2):762. PMID: 39859474
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Mol Med Rep2025 Aug;32(2):211. PMID: 40417885
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iScienceDevelopment of the FGFR4-ADC bearing the ferroptosis inducer sulfasalazine for hepatocellular carcinoma. [Abstract]2025 Oct 6;28(11):113718. PMID: 41210985
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J VirolEquine arteritis virus Nsp10 promotes MAVS proteasomal degradation via E3 ligases Smurf1/MARCH5. [Abstract]2026 Jun 23;100(6):e0206125. PMID: 42153738
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J VirolThe mRNA and protein of IL-8 oppositely regulate PRRSV replication via nucleoprotein and 14-3-3γ. [Abstract]2025 Aug 19;99(8):e0065525. PMID: 40704900
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BMC Cancer2025 Jul 17;25(1):1182. PMID: 40676553
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Discov OncolThe role of RAB GTPases in predicting prognosis and therapy response in pancreatic cancer. [Abstract]2025 Jul 21;16(1):1383. PMID: 40690079
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J Mol HistolAstragaloside II suppresses colorectal cancer progression by upregulating LGALS4 expression. [Abstract]2026 Jun 29;57(4):199. PMID: 42319491
Storage
-20°C; 2 years.
Components
| Components | HY-K1057-1 mL | HY-K1057-5 mL |
|---|---|---|
| Puromycin, Sterile (10 mg/mL) | 1 mL | 5 mL |