TR-1003
Polybrene Infection / Transfection Reagent
A highly efficient method of gene transfer into mammalian cells leveraging infection with retroviral vectors.
Sinônimo(s):
1,5-Dimethyl-1,5-diazaundecamethylene polymethobromide, hexamethrine bromide, retroviral infection
Faça loginpara ver os preços organizacionais e de contrato
About This Item
Código UNSPSC:
41122100
eCl@ss:
32160801
NACRES:
NA.75
Produtos recomendados
Nível de qualidade
forma
liquid
fabricante/nome comercial
Specialty Media
técnica(s)
transfection: suitable
Categorias relacionadas
Aplicação
Polybrene is a cationic polymer that can greatly enhance the efficiency of the retroviral or lentiviral infection to the mammalian cells. It acts to neutralize the charge repulsion between virions and the cell surface, thereby increasing infection efficiency. The efficiency of retroviral infection is enhanced significantly, 100 to 1,000 fold in some cells, by including polybrene during the infection. Polybrene with a DMSO shock is also used to mediate DNA transfer into a variety of cell types, such as CHO, chicken embryo fibroblasts, NINH-3T3 and Myeloid cells.
Protocol: Retroviral Infection
Recombinant retroviral stocks are prepared by adding 5mls of growth medium with 5% serum to a near confluent monolayer of transfected retroviral packaging cells in a 100mm plate. After 24 hours the medium is removed and filtered through a 0.45um filter.
Cells to be infected with this recombinant retroviral stock are plated at 500,000 cells per 100mm plate in 10mls of complete medium.
24 hours later, remove the growth medium from the cells. Infect cells with 2mls of the viral supernatant (or a dilution of the virus stock into 2mls) in the presence of 5ug to 10ug of polybrene per ml (final concentration). Incubate cells for 3 to 6 hours at 37°C.
Add 8mls of complete medium. Three days after infection, split the cells 1:5 into selection medium.
Protocol: Transfection
Plate cells at approximately 50% confluence in complete growth medium.
18 to 24 hours post plating, prepare the DNA-Medium-Polybrene solution, immediately before using as follows:
Note: Each component must be added in the proper sequence.
1st: Complete growth medium (2mls for a 60mm plate and 3mls for
a 100mm plate) warmed to 37°C.
2nd: Plasmid DNA, 10ng to 10ug. Gently mix.
3rd: Polybrene to a final concentration of 5ug to 10ug per ml. Gently mix
Remove medium from plate and add DNA-Medium-Polybrene solution to cells. Incubate cells at 37°C for 6 to 20 hours with occasional gentle rocking approximately every 1.5 hours for the first 6 hours.
Remove DNA-Medium-Polybrene solution and gently overlay cells with DMSO shock solution (15% DMSO in 1X HBSS: Specialty Media catalog #S-051-D) 3mls per 60mm dish and 4mls per 100mm plate. Manually rock the dish for 10 seconds to evenly distribute the solution, and then incubate the cells for exactly 4 minutes at 37°C.
Immediately remove the DMSO shock solution and gently rinse the cells twice with complete growth medium, 5mls per wash per 60mm dish, 10mls per wash per 100mm dish
Add complete growth medium to the cells.
For Stable transformants, remove the growth media and split the cells 1:5 into selection medium.
For transient expression, remove the growth medium and add fresh growth medium. Harvest cells and/or medium after 24 to 72 hours.
Reagent is supplied filtered through 0.2μm membranes and hydrated with sterile H2O.
Protocol: Retroviral Infection
Recombinant retroviral stocks are prepared by adding 5mls of growth medium with 5% serum to a near confluent monolayer of transfected retroviral packaging cells in a 100mm plate. After 24 hours the medium is removed and filtered through a 0.45um filter.
Cells to be infected with this recombinant retroviral stock are plated at 500,000 cells per 100mm plate in 10mls of complete medium.
24 hours later, remove the growth medium from the cells. Infect cells with 2mls of the viral supernatant (or a dilution of the virus stock into 2mls) in the presence of 5ug to 10ug of polybrene per ml (final concentration). Incubate cells for 3 to 6 hours at 37°C.
Add 8mls of complete medium. Three days after infection, split the cells 1:5 into selection medium.
Protocol: Transfection
Plate cells at approximately 50% confluence in complete growth medium.
18 to 24 hours post plating, prepare the DNA-Medium-Polybrene solution, immediately before using as follows:
Note: Each component must be added in the proper sequence.
1st: Complete growth medium (2mls for a 60mm plate and 3mls for
a 100mm plate) warmed to 37°C.
2nd: Plasmid DNA, 10ng to 10ug. Gently mix.
3rd: Polybrene to a final concentration of 5ug to 10ug per ml. Gently mix
Remove medium from plate and add DNA-Medium-Polybrene solution to cells. Incubate cells at 37°C for 6 to 20 hours with occasional gentle rocking approximately every 1.5 hours for the first 6 hours.
Remove DNA-Medium-Polybrene solution and gently overlay cells with DMSO shock solution (15% DMSO in 1X HBSS: Specialty Media catalog #S-051-D) 3mls per 60mm dish and 4mls per 100mm plate. Manually rock the dish for 10 seconds to evenly distribute the solution, and then incubate the cells for exactly 4 minutes at 37°C.
Immediately remove the DMSO shock solution and gently rinse the cells twice with complete growth medium, 5mls per wash per 60mm dish, 10mls per wash per 100mm dish
Add complete growth medium to the cells.
For Stable transformants, remove the growth media and split the cells 1:5 into selection medium.
For transient expression, remove the growth medium and add fresh growth medium. Harvest cells and/or medium after 24 to 72 hours.
Reagent is supplied filtered through 0.2μm membranes and hydrated with sterile H2O.
forma física
10mg polybrene per mL sterile Ultra Pure water.
Armazenamento e estabilidade
Store at -20°C upto 2 years.
Exoneração de responsabilidade
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
Código de classe de armazenamento
12 - Non Combustible Liquids
Classe de risco de água (WGK)
WGK 2
Ponto de fulgor (°F)
Not applicable
Ponto de fulgor (°C)
Not applicable
Certificados de análise (COA)
Busque Certificados de análise (COA) digitando o Número do Lote do produto. Os números de lote e remessa podem ser encontrados no rótulo de um produto após a palavra “Lot” ou “Batch”.
Já possui este produto?
Encontre a documentação dos produtos que você adquiriu recentemente na biblioteca de documentos.
Os clientes também visualizaram
Margaret M McDaniel et al.
Cell reports, 31(5), 107604-107604 (2020-05-07)
Inflammasome activation leads to pyroptotic cell death, thereby eliminating the replicative niche of virulent pathogens. Although inflammasome-associated cytokines IL-1β and IL-18 have an established role in T cell function, whether inflammasome activation in dendritic cells (DCs) is critical for T cell priming
Nam Ji Sung et al.
Bioscience reports, 40(3) (2020-03-07)
Docosahexaenoic acid (DHA) is an omega-3 fatty acid abundant in fish oils. It is known to have an inhibitory effect on various diseases such as inflammation, diabetes, and cancer. Epithelial-to-mesenchymal transition (EMT) is a process that epithelial cells gain migratory
Enhancement and inhibition of avian sarcoma viruses by polycations and polyanions.
K Toyoshima et al.
Virology, 38(3), 414-426 (1969-07-01)
W G Chaney et al.
Somatic cell and molecular genetics, 12(3), 237-244 (1986-05-01)
High-frequency transfection of CHO cells has been achieved for several plasmids, a cosmid library, and genomic DNA using Polybrene and dimethyl sulfoxide. All plasmid transfectants examined were stable and exhibited plasmid sequences in genomic DNA. The method is simple, reproducible
Marta Llaurado Fernandez et al.
Gynecologic oncology, 157(1), 12-20 (2020-01-20)
Low-grade serous ovarian carcinomas (LGSC) are frequently ER/PR positive, though the mechanisms by which ER/PR regulate prognosis or anti-estrogen treatment efficacy are poorly understood. We studied ER/PR expression in LGSC tumors and cell lines to evaluate patient outcomes and cellular
Artigos
High titer lentiviral particles for LC3 variants used for live cell analysis of cellular autophagy.
SeqPlex™-I WTA kit amplifies RNA for NGS, enabling genomic studies from limited samples.
High titer lentiviral particles including beta-actin, alpha-tubulin and vimentin used for live cell analysis of cytoskeleton structure proteins.
Protocolos
Stem cell reprogramming protocols to generate human induced pluripotent stem cells (iPSCs) including viral and non-viral RNA based methods.
Conteúdo relacionado
KOD One™ PCR Master Mix overview for ultra-fast PCR with high specificity, fidelity, and yield
Nossa equipe de cientistas tem experiência em todas as áreas de pesquisa, incluindo Life Sciences, ciência de materiais, síntese química, cromatografia, química analítica e muitas outras.
Entre em contato com a assistência técnica