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Polybrene

Polybrene, also known as hexadimethrine bromide, is a cationic polymer commonly used as a transfection agent to enhance the efficiency of virus-mediated gene delivery.
It works by neutralizing the negative charge on cell membranes, allowing for improved uptake of viral particles.
Polybrene is widely utilized in cell biology research, particularly in the fields of molecular biology and virology, to facilitate the introduction of genetic material into target cells.
Researchers can leverage PubCompare.ai's AI-powered platform to quickly identify the most reliable and optimised protocols from published literature, preprints, and patents, enabling them to enhance the reproducibility and accuracy of their Polybrene-based experiments.

Most cited protocols related to «Polybrene»

1
Efficient Transduction and Selection Protocols
Cells were transduced at a MOI between 0.5 and 1. Viruses and cells were incubated overnight in D-MEM media (Invitrogen) containing 6 µg/ml of polybrene (Sigma) in a final volume of 950 µl for 6-well plates, 5 ml for 10 cm dishes and 11 ml for 15 cm dishes. The next day, the viruses were removed, the cells were rinsed twice with PBS and fresh media was added. For primary cells, a second round of transduction was done. Drug selection was added at 48 h post-transduction. The following concentrations were used: blasticidin: 2.5 µg/ml for WI38, HCA2, BJ cells and 5 µg/ml for the other cells, hygromycin: 100 µg/ml for WI38, HCA2, BJ cells, 200 µg/ml for U2OS cells and 300 µg/ml for HT1080 cells, neomycin: 300 µg/ml for WI38, HCA2, BJ cells and 800 µg/ml for the other cells, puromycin: 0.5 µg/ml for HeLa cells and 2.0 µg/ml for other cells, zeocin: 400 µg/ml for HT1080 cells and 200 µg/ml for other cells. Induction of the cDNA/shRNA/miRNA was typically done by addition of doxycycline at a final concentration of 1.0 µg/ml for 48 h (cDNA) or 96 h (shRNA/miRNA) unless indicated otherwise.
Campeau E., Ruhl V.E., Rodier F., Smith C.L., Rahmberg B.L., Fuss J.O., Campisi J., Yaswen P., Cooper P.K, & Kaufman P.D. (2009). A Versatile Viral System for Expression and Depletion of Proteins in Mammalian Cells. PLoS ONE, 4(8), e6529.
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Publication 2009
Cells DNA, Complementary Doxycycline HeLa Cells hygromycin A Hyperostosis, Diffuse Idiopathic Skeletal MicroRNAs Neomycin Pharmaceutical Preparations Polybrene Puromycin Short Hairpin RNA Virus Zeocin
2
Generating human iPSCs from fibroblasts

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Publication 2008
Cells Doxycycline Feeder Cell Layers Fibroblasts Gelatins Homo sapiens Human Embryonic Stem Cells Human Induced Pluripotent Stem Cells Hyperostosis, Diffuse Idiopathic Skeletal Induced Pluripotent Stem Cells Infection Lentivirus Polybrene Transfection Trypsin Virus
3
Establishing Doxycycline-Inducible KSHV Models

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Publication 2011
antibiotic G 418 Antibiotics Cells Culture Media Doxycycline Germ Cells hygromycin A Hyperostosis, Diffuse Idiopathic Skeletal Polybrene Puromycin Retroviridae Viral Genome Virus
4
Monocyte-Derived Dendritic Cell Infection
Monocytes were isolated and incubated with GM-CSF and IL-4 to induce dendritic cell differentiation. Pseudotyped viruses and virus-like particles were produced by transient transfection of 293FT cells using TransIT-293 (Mirus). Infections were performed by incubating 105 MDDCs in 96 well U bottom plates in the presence of 8 µg/ml polybrene. Cell surface staining of activation markers was performed 48h after infection. shRNA vectors carrying GFP were transduced into fresh monocytes together with SIVVLP(G) and dendritic cell differentiation was induced. More than 90% of cells were routinely transduced and cells were challenged at day 4 with HDVIRESRFP(G) or other control PAMPs.
Manel N., Hogstad B., Wang Y., Levy D.E., Unutmaz D, & Littman D.R. (2010). A cryptic sensor for HIV-1 activates antiviral innate immunity in dendritic cells. Nature, 467(7312), 214-217.
Publication 2010
Cells Cloning Vectors Dendrites Dendritic Cells Differentiations, Cell Granulocyte-Macrophage Colony-Stimulating Factor Infection Macular Edema, Cystoid Monocytes Pathogen-Associated Molecular Pattern Molecules Polybrene Pseudotyped Viruses Short Hairpin RNA Transfection Transients Virion
5
Induction of Induced Pluripotent Stem Cells
Lentiviruses were produced using a five plasmid transfection system in 293T packaging cells as previously described [17 (link)]. Supernatants were collected every 12 hours during two consecutive days starting 48 hours after transfection and viral particles were concentrated by centrifugation at 16,500 rpm for 1.5 hours at 4°C. Approximately 100,000 fibroblasts were seeded on plastic in 35-mm culture plates and infected with 15 μl of concentrated virus in the presence of polybrene (5 μg/ml). The media was replaced after 16 hours with mouse ES cell media (DMEM supplemented with 15% FBS, L-glutamine, penicillin/streptomycin, nonessential amino acids, β-mercaptoethanol and 1000 U/ml LIF) and changed every 2–3 days. Doxycycline (Sigma-Aldrich, St. Louis, http://www.sigmaaldrich.com) was added at a final concentration of 1 μg/ml, where indicated, and removed at day 10 post-infection. iPS colonies were picked 20 to 25 days post-infection based on morphology and GFP expression and expanded by plating on Mitomycin C treated MEFs in ES cell media.
Sommer C.A., Stadtfeld M., Murphy G.J., Hochedlinger K., Kotton D.N, & Mostoslavsky G. (2009). iPS Cell Generation Using a Single Lentiviral Stem Cell Cassette. Stem cells (Dayton, Ohio), 27(3), 543-549.
Publication 2009
2-Mercaptoethanol Amino Acids Centrifugation Doxycycline Embryonic Stem Cells Fibroblasts Glutamine HEK293 Cells Infection Lentivirus Mitomycin Mus Penicillins Plasmids Polybrene Streptomycin Transfection Virion Virus

Most recents protocols related to «Polybrene»

1
Lentiviral Transduction of Myeloma Cells
Transient transfections to HEK 293 T cells were performed using polyethyleneimine (PEI) (Polysciences, Warrington, PA, USA) in the OPTI-MEM medium (Life Technologies, Carlsbad, CA, USA) with a ratio of 1:4 to 1:6 of DNA: PEI.
Viral particles were produced by HEK 293 T cells in a 10 cm dish transfected with 4 μg pMD2.G and 6 μg psPAX2 packaging plasmids (Addgene, Watertown, MA, USA), together with 8 μg lentiviral expressing vectors encoding target genes, including pITA -CBX3-N/C-flag, pLKO.1 or hU6-MSC-Ubiquitin-eGFP vector encoding shRNAs targeting interested genes as listed in the Supplementary Information. Supernatant carrying the viral particles was harvested 35 h and 60 h after transfection and concentrated to 100× volume by Poly (ethylene glycol) 8000 (Sigma-Aldrich, St. Louis, MS, USA).
For viral infection, 1 × 106 myeloma cells were seeded in 1 ml new complete media for 6 h and then added 50 μl viral concentration and 8 μg/mL polybrene, and cells were spin at 800 × g for 45 min at 20 °C. In total, 12 h after spinfection, the medium was changed and cells were cultured for another 48 h until further management.
Li X., Wang S., Xie Y., Jiang H., Guo J., Wang Y., Peng Z., Hu M., Wang M., Wang J., Li Q., Wang Y, & Liu Z. (2023). Deacetylation induced nuclear condensation of HP1γ promotes multiple myeloma drug resistance. Nature Communications, 14, 1290.
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Publication 2023
Cells Cloning Vectors G-800 Genes HEK293 Cells Hyperostosis, Diffuse Idiopathic Skeletal Multiple Myeloma Plasmids Polybrene polyethylene glycol 8000 Polyethyleneimine Short Hairpin RNA Transfection Transients Ubiquitin Virion Virus Diseases
2
Lentiviral Transduction of Activated Mouse CD4+ T Cells
On day 1, HEK293T cells were transfected with 1 μg of vector plus 0.5 μg of pCL-Eco plasmid (Naviaux et al., 1996 (link)) using X-tremeGene 9 DNA transfection reagent (Roche). On day 2, the media was exchanged with 1 ml fresh T cell media. CD4+ T cells were isolated from the spleens of B6 mice using an EasySep Mouse Naive CD4+ T cell isolation kit (Stemcell Technologies) and cultured in 96-well plates coated with anti-CD3 antibody (10 μg/ml; BD Biosciences) at a density of 1–2 × 106 cells per well in 0.1 ml of T cell media with soluble anti-CD28 antibody (10 μg/ml; BD Biosciences). On day 3, the activated T cells were transferred to 24-well plates at a density of 1–2 × 106 cells per well, washed, and then resuspended in virus-containing supernatants from the HEK293T cell cultures. Polybrene was added to a final concentration of 8 μg/ml. The cell–virus mixture was then centrifuged at 3,500 rpm for 90 min at 32°C to achieve transduction. Following transduction, cells were cultured in 96-well plates coated with anti-CD3 antibody (1 μg/ml) at a density of 5 × 105 cells per well in T cell media with soluble anti-CD28 antibody (1 μg/ml). On the following day, cells were washed and cultured in fresh 96-well plates in T cell media with IL-2 (20 ng/ml) until ready for further analysis.
Seth A., Yokokura Y., Choi J.Y., Shyer J.A., Vidyarthi A, & Craft J. (2023). AP-1–independent NFAT signaling maintains follicular T cell function in infection and autoimmunity. The Journal of Experimental Medicine, 220(5), e20211110.
Publication 2023
Antibodies, Anti-Idiotypic CD4 Positive T Lymphocytes Cell Culture Techniques Cells Cell Separation Cloning Vectors Mus Plasmids Polybrene Stem Cells T-Lymphocyte Transfection Virus
3
PVT1 Modulation in HNSCC Cell Lines
The American Type Culture Collection (ATCC, Manassas, VA, USA) provided the human HNSCC cell lines SCC15. HN6 was obtained from the Central Laboratory of Peking University School and Hospital of Stomatology. Dulbecco’s modified Eagle’s medium (DMEM) with 10% fetal bovine serum (FBS) and 1% antibiotics was used to culture the cells at 37 °C, 5% CO2 in a humidified incubator. Cells were transfected with miRNA mimics (Integrated Biotech Solutions Co., Ltd) or siRNA (Tsingke Biotechnology, Beijing, China) using the Lipo8000TM Transfection Reagent (Beyotime, Shanghai, China) following the manufacturer’s instructions. The lentiviruses used for PVT1 KD or overexpression were from Integrated Biotech Solutions Co., Ltd (Shanghai, China). Briefly, the scramble control or the specific PVT1 KD (shPVT1) and overexpression (pQLL-PVT1) lentiviral plasmids were cotransfected into HEK293T cells with two helper plasmids psPAX2 (Addgene, Watertown, MA, USA Cat#12260) and pMD2.G (Addgene, Cat#12259). Viral supernatants were harvested for cell infection 72 h after transfection. Cells were infected with lentiviruses in the presence of polybrene (Sigma-Aldrich, Shanghai, China, Cat#H9268), selected with puromycin (Sigma-Aldrich, Cat#P9620) and expanded before being used for subsequent assays.
Qin Z., Zhang W., Liu S., Wang Y., Peng X, & Jia L. (2023). PVT1 inhibition stimulates anti-tumor immunity, prevents metastasis, and depletes cancer stem cells in squamous cell carcinoma. Cell Death & Disease, 14(3), 187.
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Publication 2023
Antibiotics Biological Assay Cell Lines Cells Eagle Fetal Bovine Serum Homo sapiens Infection Lentivirus MicroRNAs Plasmids Polybrene Puromycin PVT1 long-non-coding RNA, human RNA, Small Interfering Squamous Cell Carcinoma of the Head and Neck Transfection
4
Lentiviral Transduction of OE19 Cells with inducible dnFOS
pINDUCER20-GFP-AFOS (ADS5006, Britton et al., 2017 (link)) was packaged into lentivirus and OE19 cells were transduced with lentivirus as previously described (Tiscornia et al., 2006 (link)). Briefly, 3 × 106 HEK293T cells were transfected with 2.25 μg psPAX2 (Addgene, 12260), 1.5 μg pMD2.G (Addgene, 12259), and 3 μg pINDUCER20-GFP-AFOS using PolyFect (Qiagen, 301107). Media was collected at 48 and 72 hr post-transfection and viral particles were precipitated using PEG-it Solution (System Biosciences, LV810A-1). To transduce, cells were treated with virus (Multiplicity of Infection (MOI) 0.5–1.0) and 5 μg/ml Polybrene (EMD Millipore, TR-1003). Polyclonal cells were selected for 2 weeks in 250 μg/ml G418 (Thermo Fisher Scientific, 10131027). dnFOS (Olive et al., 1997 (link)) was induced with 1 μg/ml doxycycline.
Ahmed I., Yang S.H., Ogden S., Zhang W., Li Y, & Sharrocks A.D. (2023). eRNA profiling uncovers the enhancer landscape of oesophageal adenocarcinoma and reveals new deregulated pathways. eLife, 12, e80840.
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Publication 2023
antibiotic G 418 Cells Doxycycline Infection Lentivirus Olea europaea Polybrene Transfection Virion Virus
5
Lentiviral Transduction of Cancer Cell Lines
HEK293T cells were cultured until 60% confluence in 6 cm plates and cotransfected with 2 μg of the appropriate plasmids and 2.5 μg of helper plasmids (1.5 μg psPAX2 and 1 μg pmD2.G) using Lipofectamine 2000 (Invitrogen) according to the manufacturer’s instructions. The supernatants containing viral particles were collected 48 h and 72 h later and filtered through 0.45 μm polyvinylidene fluoride (PVDF) membranes. MDA-MB-231, MCF-7, and 4T1 cells were transduced with viral supernatants containing 5 μg/mL polybrene, and the medium was replaced 48 h later. The stably transduced cells were selected by culturing in the presence of 2 μg/mL puromycin for at least 7 days. Subsequently, the cells were harvested for Western blotting.
Zhang J., Guo F., Li C., Wang Y., Wang J., Sun F., Zhou Y., Ma F., Zhang B, & Qian H. (2023). Loss of TTC17 promotes breast cancer metastasis through RAP1/CDC42 signaling and sensitizes it to rapamycin and paclitaxel. Cell & Bioscience, 13, 50.
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Publication 2023
Cells lipofectamine 2000 Plasmids Polybrene polyvinylidene fluoride Puromycin Tissue, Membrane Virion

Top products related to «Polybrene»

1
Polybrene by Merck Group
Sourced in United States, Germany, China, Sao Tome and Principe, United Kingdom, Macao, Canada, France, Japan, Switzerland, Italy, Australia, Netherlands, Morocco, Israel, Ireland, Belgium, Denmark, Norway
Polybrene is a cationic polymer used as a transfection reagent in cell biology research. It facilitates the introduction of genetic material into cells by enhancing the efficiency of DNA or RNA uptake.
2
Lipofectamine 2000 by Thermo Fisher Scientific
Sourced in United States, China, Germany, United Kingdom, Canada, Japan, France, Italy, Switzerland, Australia, Spain, Belgium, Denmark, Singapore, India, Netherlands, Sweden, New Zealand, Portugal, Poland, Israel, Lithuania, Hong Kong, Argentina, Ireland, Austria, Czechia, Cameroon, Taiwan, Province of China, Morocco
Lipofectamine 2000 is a cationic lipid-based transfection reagent designed for efficient and reliable delivery of nucleic acids, such as plasmid DNA and small interfering RNA (siRNA), into a wide range of eukaryotic cell types. It facilitates the formation of complexes between the nucleic acid and the lipid components, which can then be introduced into cells to enable gene expression or gene silencing studies.
3
Puromycin by Merck Group
Sourced in United States, Germany, China, Sao Tome and Principe, United Kingdom, Macao, Canada, Japan, Italy, Switzerland, France, Israel, Australia, Spain, Belgium, Morocco, Sweden
Puromycin is a laboratory product manufactured by Merck Group. It functions as an antibiotic that inhibits protein synthesis in eukaryotic cells.
4
Pspax2 by Addgene
Sourced in United States, Switzerland, China, Japan, Germany, United Kingdom, France, Canada
PsPAX2 is a packaging plasmid used for the production of lentiviral particles. It contains the necessary genes for lentiviral packaging, but does not contain the viral genome. PsPAX2 is commonly used in lentiviral production workflows.
5
Pmd2 g by Addgene
Sourced in United States, Switzerland, China, Germany, France, Japan, United Kingdom, Canada
The PMD2.G is a lab equipment product. It is a plasmid that can be used for various research applications.
6
Puromycin by Thermo Fisher Scientific
Sourced in United States, Germany, China, United Kingdom, France, Canada, Japan, Australia, Switzerland, Belgium, Italy, Macao
Puromycin is a laboratory reagent used for selection of mammalian cells expressing a puromycin resistance gene. It acts as an antibiotic that inhibits protein synthesis, leading to cell death in cells that do not express the resistance gene.
7
Polybrene by Santa Cruz Biotechnology
Sourced in United States, Germany, Canada, China
Polybrene is a cationic polymer that helps increase the efficiency of viral transduction in cell culture experiments. It functions by facilitating the interaction between the virus and the target cells, thereby enhancing viral entry and subsequent gene delivery.
8
Lipofectamine 3000 by Thermo Fisher Scientific
Sourced in United States, China, Germany, Japan, United Kingdom, France, Canada, Italy, Australia, Switzerland, Denmark, Spain, Singapore, Belgium, Lithuania, Israel, Sweden, Austria, Moldova, Republic of, Greece, Azerbaijan, Finland
Lipofectamine 3000 is a transfection reagent used for the efficient delivery of nucleic acids, such as plasmid DNA, siRNA, and mRNA, into a variety of mammalian cell types. It facilitates the entry of these molecules into the cells, enabling their expression or silencing.
9
Fetal bovine serum (fbs) by Thermo Fisher Scientific
Sourced in United States, China, United Kingdom, Germany, Australia, Japan, Canada, Italy, France, Switzerland, New Zealand, Brazil, Belgium, India, Spain, Israel, Austria, Poland, Ireland, Sweden, Macao, Netherlands, Denmark, Cameroon, Singapore, Portugal, Argentina, Holy See (Vatican City State), Morocco, Uruguay, Mexico, Thailand, Sao Tome and Principe, Hungary, Panama, Hong Kong, Norway, United Arab Emirates, Czechia, Russian Federation, Chile, Moldova, Republic of, Gabon, Palestine, State of, Saudi Arabia, Senegal
Fetal Bovine Serum (FBS) is a cell culture supplement derived from the blood of bovine fetuses. FBS provides a source of proteins, growth factors, and other components that support the growth and maintenance of various cell types in in vitro cell culture applications.
10
Lenti x concentrator by Takara Bio
Sourced in United States, Japan, Germany, Canada, France
The Lenti-X concentrator is a device used to concentrate lentiviral particles. It functions by facilitating the concentration of lentiviral particles through a centrifugation process.

More about "Polybrene"

Polybrene, also known as hexadimethrine bromide, is a cationic polymer that enhances the efficiency of virus-mediated gene delivery.
It works by neutralizing the negative charge on cell membranes, allowing for improved uptake of viral particles.
Polybrene is widely used in cell biology research, particularly in molecular biology and virology, to facilitate the introduction of genetic material into target cells.
Researchers often combine Polybrene with other transfection agents like Lipofectamine 2000, Puromycin, PsPAX2, and PMD2.G to optimize their experiments.
Lipofectamine 3000 is another popular transfection reagent that can be used in conjunction with Polybrene.
Additionally, the addition of Fetal Bovine Serum (FBS) and the use of Lenti-X concentrator can further improve the efficiency of Polybrene-based gene delivery.
PubCompare.ai's AI-powered platform can help researchers quickly identify the most reliable and optimized protocols from published literature, preprints, and patents.
This enables them to enhance the reproducibility and accuracy of their Polybrene-based experiments, leading to more robust and reliable results.
By leveraging the insights and tools provided by PubCompare.ai, researchers can streamline their workflow and optimize their use of Polybrene, ultimately advancing their scientific discoveries.