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P9620 Sigma

Puromycin dihydrochloride

Ready Made Solution, from Streptomyces alboniger, 10 mg/mL in H2O, 0.2 μm filtered

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Description

Preparation Note

This product is concentrated at 10 mg/mL in water and is 0.2 μm filtered. It has been tested on HeLa cells for cell growth arrest and selection of cells after transfection of the pac resistance gene.

Application

Puromycin is an aminonucleoside antibiotic that is derived from Streptomyces alboniger. It allows selection for cells that contain the resistance gene puromycin N-acetyl-transferase (PAC). It has been used to study vascular smooth muscle cell viability after treatment in the rabbit model 1. Puromycin is used to produce enhanced green fluorescent protein (EGFP) transgenic piglets after somatic cell cloning and embryo transfer2.

Biochem/physiol Actions

Mode of Action: Puromycin inhibits protein synthesis by causing premature chain termination acting as an analog of the 3′-terminal end of the aminoacyl-tRNA. Puromycin also acts as a reversible inhibitor of dipeptidyl-peptidase II (serine peptidase) and cytosol alanyl aminopeptidase.

Mode of Resistance: Puromycin acetyltransferase is an effective resistance gene.

Antimicrobial Spectrum: This product is active against gram-positive microorganisms, less active against acid-fast bacilli and more weakly active against gram-negative microorganisms. Puromycin can prevent growth of bacteria, protozoa, algae and mammalian cells and acts quickly, killing 99% of cells within 2 days.

Protocols & Applications

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Puromycin dihydrochloride from <I>Streptomyces alboniger</I>

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Safety & Documentation

Protocols & Articles

Articles

Examination of Efficacy, Specificity, and Efficiency of an shRNA Lentiviral Library

Betsy Boedeker,* Gregory A. Wemhoff, Andrea Spencer, Roger Chiu, Suzanne Miller, Henry George, and Edward J. Weinstein
Betsy Boedeker, Gregory A. Wemhoff, Andrea Spencer,Roger Chiu, Suzanne Miller, Henry George and Edward J. Weinstein
LSI Volume 7 Article 1
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The MISSION® Target ID Library for Human miRNA Target Identification and Discovery

One of the main problems miRNA researchers face today is global identification of gene targets that are functionally regulated by their miRNA of interest. Unfortunately, bioinformatic predictions are...
Keywords: AGE, Amplification, Apoptosis, Cancer, Cell culture, Cloning, Degradations, Detection methods, Digestions, Electrophoresis, Environmental, Fractionation, Gel electrophoresis, Gene expression, Genomics, Immunoprecipitation, Melting, Methods, Microarray Analysis, Polymerase chain reaction, Polymerase chain reaction - quantitative, Purification, RNA immunoprecipitation, Reductions, Sequences, Sequencing, Support, Transfection, Western blot, transformation

Protocols

Protocol for Transducing Mouse Embryonic Fibroblasts (MEF) using MISSION® ExpressMag™ Super Magnetic Kit®

0.1 Seed 15k MEF per well into 96-well plate. Utilize a final volume of 200 µl of media per well. (Note: for best data interpretation always include an untransduced well that is treated with puromyci...

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Bio Brief: Puromycin [VIDEO]

Puromycin dihydrochloride Ready Made Solution is supplied as a ready-to-use, sterile-filtered solution at a 10 mg/ml concentration. Puromycin is an aminonucleoside antibiotic produced by Streptomyces...
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Genome-wide Screen for miRNA Targets Using the MISSION® Target ID Library [VIDEO]

The MISSION Target ID Library is designed to assist in discovery and identification of microRNA (miRNA) targets. The MISSION Target ID Library is a plasmid-based, genome-wide cDNA library cloned into...
Keywords: PAGE, Sequencing

Peer-Reviewed Papers

References

Set your institution to view full text papers.

1. Differential Effect of the Protein Synthesis Inhibitors Puromycin and Cycloheximide on Vascular Smooth Muscle Cell Viability Valerie Croons, Wim Martinet, et al. J. Pharmacol. Exp. Ther. 325, 824-832, (2008)

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2. A novel method for the production of transgenic cloned pigs: electroporation-mediated gene transfer to non-cultured cells and subsequent selection with puromycin. Watanabe S, Iwamoto M, et. al. Biol. Reprod. 72, 309-315, (2005)

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Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Takahashi, K. and Yamanaka S. Cell 126, 663-676, (2006)

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Design and application of a cytokine-receptor-based interaction trap. Eyckerman, S., et al. Nat. Cell Biol. 3, 1114-1116, (2001)

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Tabuchi, I. Biochem. Biophys. Res. Commun. 23, 2-5, (2003)

ERK- And Akt-dependent Neuroprotection By Erythropoietin (EPO) Against Glyoxal-AGEs Via Modulation Of Bcl-xL, Bax, And BAD. Shen, J., et al. Invest. Ophthalmol. Vis. Sci. 51, 35-46, (2010)

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Rapamycin Activates Autophagy And Improves Myelination In Explant Cultures From Neuropathic Mice. Rangaraju, S., et al. J. Neurosci. 30, 11388-97, (2010)

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Selective suicide of cross-presenting CD8+ dendritic cells by cytochrome c injection shows functional heterogeneity within this subset. Ming Lee Lin et al Proc. Natl. Acad. Sci. U. S. A. 105, 3029-34, (2008)

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Multiple cellular mechanisms related to cyclin A1 in prostate cancer invasion and metastasis. Wegiel B J. Natl. Cancer Inst. 100(14), 1022-36, (2008)

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Cyclin-dependent kinase 5 is a calmodulin-binding protein that associates with puromycin-sensitive aminopeptidase in the nucleus of Dictyostelium. Huber RJ, Catalano A, and O'Day DH Biochim. Biophys. Acta 1833(1), 11-20, (2013)

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Genome-wide search for novel human uORFs and N-terminal protein extensions using ribosomal footprinting. Fritsch C, Herrmann A, Nothnagel M, et al. Genome Res. 22(11), 2208-18, (2012)

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Application of mRNA display for in vitro selection of DNA-binding transcription factor complexes. Tateyama S and Yanagawa H Methods Mol. Biol. 977, 95-109, (2013)

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Measurement of protein synthesis: in vitro comparison of (68)Ga-DOTA-puromycin, [ (3)H]tyrosine, and 2-fluoro-[ (3)H]tyrosine. Eigner S, Beckford Vera DR, Fellner M, et al. Recent Results Cancer Res. 194, 269-83, (2013)

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Scandium-44: benefits of a long-lived PET radionuclide available from the (44)Ti/(44)Sc generator system. Roesch F Curr. Radiopharm. 5(3), 187-201, (2012)

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Modulation of ER stress and apoptosis by endoplasmic reticulum calcium leak via translocon during unfolded protein response: involvement of GRP78. Hammadi M, Oulidi A, Gackière F, et al. FASEB J. 27(4), 1600-9, (2013)

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Improvement of a puromycin-linker to extend the selection target varieties in cDNA display method. Ueno S, Kimura S, Ichiki T, et al. J. Biotechnol. 162(2-3), 299-302, (2012)

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Haematopoietic stem cells require a highly regulated protein synthesis rate. Signer RA, Magee JA, Salic A, et al. Nature 509(7498), 49-54, (2014)

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Elucidating the regulon of multidrug resistance regulator RarA in Klebsiella pneumoniae. De Majumdar S, Veleba M, Finn S, et al. Antimicrob. Agents Chemother. 57(4), 1603-9, (2013)

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TDP2/TTRAP is the major 5'-tyrosyl DNA phosphodiesterase activity in vertebrate cells and is critical for cellular resistance to topoisomerase II-induced DNA damage. Zhihong Zeng et al J. Biol. Chem. 286, 403-9, (2011)

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Tumour microenvironments induce expression of urokinase plasminogen activator receptor (uPAR) and concomitant activation of gelatinolytic enzymes. Magnussen S, Hadler-Olsen E, Latysheva N, et al. PLoS ONE 9(8), e105929, (2014)

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EGFR-mediated chromatin condensation protects KRAS-mutant cancer cells against ionizing radiation. Wang M, Kern AM, Hülskötter M, et al. Cancer Res. 74(10), 2825-34, (2014)

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Stable transfection of the diplomonad parasite Spironucleus salmonicida. Jerlström-Hultqvist J, Einarsson E, and Svärd SG Eukaryotic Cell 11(11), 1353-61, (2012)

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A novel inducible protein production system and neomycin resistance as selection marker for Methanosarcina mazei. Mondorf S, Deppenmeier U, and Welte C Archaea 2012, 973743, (2012)

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Selective desulfurization significantly expands sequence variety of 3'-peptidyl-tRNA mimics obtained by native chemical ligation. Geiermann AS and Micura R ChemBioChem. 13(12), 1742-5, (2012)

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Protein kinase PKR mediates the apoptosis induction and growth restriction phenotypes of C protein-deficient measles virus. Toth AM, Devaux P, Cattaneo R, et al. J. Virol. 83(2), 961-8, (2009)

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DEF6, a novel substrate for the Tec kinase ITK, contains a glutamine-rich aggregation-prone region and forms cytoplasmic granules that co-localize with P-bodies. Hey F, Czyzewicz N, Jones P, et al. J. Biol. Chem. 287(37), 31073-84, (2012)

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NADPH oxidase-mediated upregulation of connexin43 contributes to podocyte injury. Yan Q, Gao K, Chi Y, et al. Free Radic. Biol. Med. 53(6), 1286-97, (2012)

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Poly(A) binding protein C1 is essential for efficient L1 retrotransposition and affects L1 RNP formation. Dai L, Taylor MS, O'Donnell KA, et al. Mol. Cell. Biol. 32(21), 4323-36, (2012)

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Kaposi's sarcoma-associated herpesvirus viral interferon regulatory factor 4 targets MDM2 to deregulate the p53 tumor suppressor pathway. Hye-Ra Lee et al J. Virol. 83, 6739-47, (2009)

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Imaging translation in single cells using fluorescent microscopy. Chao JA, Yoon YJ, and Singer RH Cold Spring Harb. Perspect. Biol. 4(11), doi:10.1101/cshperspect.a012310, (2012)

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Receptor activator of NF-kappaB and podocytes: towards a function of a novel receptor-ligand pair in the survival response of podocyte injury. Liu S, Shi W, Xiao H, et al. PLoS ONE 7(7), e41331, (2012)

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Caspase-3 cleaves hnRNP K in erythroid differentiation. Naarmann-de Vries IS, Urlaub H, Ostareck DH, et al. Cell Death Dis. 4, e548, (2013)

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Regulation of Tsg101 expression by the steadiness box: a role of Tsg101-associated ligase. McDonald B Mol. Biol. Cell 19(2), 754-63, (2008)

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Glo1 genetic amplification as a potential therapeutic target in hepatocellular carcinoma. Zhang S, Liang X, Zheng X, et al. Int. J. Clin. Exp. Pathol. 7(5), 2079-90, (2014)

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Premature translational termination products are rapidly degraded substrates for MHC class I presentation. Lacsina JR, Marks OA, Liu X, et al. PLoS ONE 7(12), e51968, (2012)

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Thrombin-sensitive expression of the store operated Ca(2+) channel Orai1 in platelets. Münzer P, Tolios A, Pelzl L, et al. Biochem. Biophys. Res. Commun. 436(1), 25-30, (2013)

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The dynamics of in vitro maturation of germinal vesicle oocytes. Escrich L, Grau N, de los Santos MJ, et al. Fertil. Steril. 98(5), 1147-51, (2012)

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Deficiency of ALADIN impairs redox homeostasis in human adrenal cells and inhibits steroidogenesis. Prasad R, Metherell LA, Clark AJ, et al. Endocrinology 154(9), 3209-18, (2013)

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A pull-down method with a biotinylated bait protein prepared by cell-free translation using a puromycin linker. Mochizuki Y, Kohno F, Nishigaki K, et al. Anal. Biochem. 434(1), 93-5, (2013)

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Suppression of Aβ toxicity by puromycin-sensitive aminopeptidase is independent of its proteolytic activity. Kruppa AJ, Ott S, Chandraratna DS, et al. Biochim. Biophys. Acta 1832(12), 2115-26, (2013)

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Host proteolytic activity is necessary for infectious bursal disease virus capsid protein assembly. Irigoyen N, Castón JR, and Rodríguez JF J. Biol. Chem. 287(29), 24473-82, (2012)

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Pulsed electromagnetic field affects intrinsic and endoplasmatic reticulum apoptosis induction pathways in MonoMac6 cell line culture. Kaszuba-Zwoinska J, Chorobik P, Juszczak K, et al. J. Physiol. Pharmacol. 63(5), 537-45, (2012)

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Mechanisms of Bv8-induced biphasic hyperalgesia: increased excitatory transmitter release and expression. De Felice M, Melchiorri P, Ossipov MH, et al. Neurosci. Lett. 521(1), 40-5, (2012)

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Superinduction of leptin mRNA in mouse hypothalamic neurons. Brown RE, Wilkinson DA, Wilkinson PM, et al. Neuroreport 23(15), 900-3, (2012)

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The persistent plausibility of protein synthesis in the nucleus: process, palimpsest or pitfall? Pederson T Curr. Opin. Cell Biol. 25(4), 520-1, (2013)

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