Fugene hd transfection reagent lets you work in your cancer model. Designed for dramatically improved transfection efficiency, invitrogen lipofectamine stem transfection reagent offers a simple. Promega corporation 2800 woods hollow road madison, wi 537115399 usa toll free. Genejuice transfection reagent nonlipid based chemical transfection reagent optimized for maximum transfection efficiency, easeofuse, and minimal cytotoxicity on a wide variety of mammallian cells. Storage and stability fugene 6 reagent is shipped at room temperature. The transfection protocol for lipofectamine 3000 was developed to be easy to use while still ensuring optimum performance and. Enhanced and simple transfection protocol for a variety of cell lines. Fugene hd transfection reagent is a multicomponent reagent that forms a complex with dna, then transports the complex into animal or insect cells. Using the nanobit ppi system with glomax discover to. Xtremegene transfection reagents from roche have been used for a wide range of applications and a vast array of cell lines, primary cells and stem cells. Reproducibly high transfection rates with low methodologyinduced cytotoxic side effects are essential to attain the required effect on targeted cells when exogenous dna is transfected. Figure 1 vertiga shaker manufactured by thomson instruments co. Using the nanobit ppi system with glomax discover to detect proteinprotein interactions promega corporation.
The fugene hd protocol database is a collection of protocols that guide the transfection process for a variety of cell lines. Utilizes a protocol that does not require removal of serum or culture medium and does not require washing or changing of medium after introducing the reagentdna. Stem cells protocols complete protocol fugene hd transfection reagent. Fugene 6 transfection reagent is gentle on the cells. Olson 2000 activation of the myocyte enhancer factor2 transcription factor by calciumcalmodulindependent protein kinasestimulated binding of 1433 to histone deacetylase 5. Lipofectamine stem transfection reagent thermo fisher.
The transitlt1 reagent exhibits higher expression and lower cellular toxicity compared to other transfection reagents. Oct 25, 2012 this protocol describes an edtabased passaging procedure to be used with chemically defined e8 medium that serves as a tool for basic and translational research into human pluripotent stem cells. In the dlr assay, the activities of firefly and renilla luciferases are measured sequentially from a single sample. Hepg2 cells were transfected with a luciferase expression plasmid using the designated reagents at the manufacturers recommended reagenttodna ratio indicated beneath each bar. Generation of mouseinduced pluripotent stem cells with. Fugene hd transfection reagent quick protocol fb112pdf. Fugener hd transfection reagent technical manual tm328. Fugener hd transfection reagent quick protocol card, fb112. Transfection efficiency and protein expression in various cell lines. Human es h9 cells were transfected with lipofectamine stem reagent or fugene hd reagent according to. Citation list of established cells transfected with fugene 6 transfection reagent 10t12 timothy a. The reagent is stable thro ugh the expiration date printed on the label when stored under these conditions. This protocol describes an edtabased passaging procedure to be used with chemically defined e8 medium that serves as a tool for basic and translational research into.
The fugene hd transfection reagent protocol database has protocols for transfecting your cell line. Fugene hd has an exclusive ability to perform transfection in 100% serum, thus closely mimicking transfection in in vivo conditions. Transfection of sf9 cells in suspension december, 2008 version 1. The database includes over 25 cancer models and provides transfection protocols specific for each cell line. Lipofectamine 3000 reagent provided higher gfp transfection efficiency than lipofectamine 2000 and fugene hd reagents for all five cell lines. Shop online for a wide selection of promega fugene hd transfection reagent fugene hd is a novel, nonliposomal reagent for transfecting dna into a variety of cell lines with. The fugene hd transfection reagent is a nonliposomal formulation designed to transfect dna into a wide variety of cell lines with high efficiency and low toxicity. Use the fugene hd protocol database to see if transfection conditions for your cell line are available. Recommended transfection protocol for initial transfection experiment and transfection optimization we suggest using 96well plate format. Fugene hd transfection reagent is a proprietary, nonliposomal, low toxicity formulation designed for high efficiency delivery of dna into a broad range of cell lines. The use of fugene hd does not require culture medium change or serum removal during transfection experiment. Transfection is the process of deliberately introducing naked or purified nucleic acids into eukaryotic cells. Fugene 6 transfection reagent university of pittsburgh.
Citation list of established cells transfected with fugene 6. Cells that have incorporated the foreign dna are called transfectants. Fugene 6 transfection reagent is a nonliposomal formulation designed to transfect plasmid dna into a wide variety of cell lines with high efficiency and low toxicity. The protocol does not require removal of serum or culture medium and does not require washing or changing of medium after introducing the reagentdna.
Citation list of established cells transfected with fugene. Transfection of sf9 cells in suspension nih common fund. The transfection protocol for lipofectamine 3000 was developed to be easy to use while still ensuring optimum performance and reliability in a wide panel of cell lines. Vortex and incubate the mixture at room temperature for 15 min before adding to the well.
Additionally, low cell numbers can be transfected in 96well plates. Fugene 6 transfection reagent has been commercially available since 1997. Newer reagents such as fugene hd involve a single addition of. Adherent cells can be trypsinized and transfected by the dna. Fugene hd transfection reagent also provides quicker results in less time with its simple, consistent, twostep protocol applicable across a wide range of cell types. Fugene hd transfection reagent is a nonliposomal formulation designed to transfect dna into a wide.
This protocol is used to generate baculoviruses containing gpcr genes of interest for expression in sf9 cells. We routinely use fugene hd to introduce plasmids encoding tagged lipidbinding domains into raw264. Using the nanobit ppi system with glomax discover to detect. Fugene hd transfection reagent protocol promega corporation. Fugene hd transfection reagent roche applied science. Promega fugene hd transfection reagent fugene hd is a novel, nonliposomal reagent for transfecting dna into a variety of cell lines with high efficiency and low toxicity. In animal cells, transfection is the preferred term as. During development of each reagent, good transfection efficiency was verified in a variety of cells as indicated in the product manual. Fugene 6 reagent complex prior to plating, making it a strong candidate for high throughput applications. Hepg2 cells were transfected with a luciferase expression plasmid using the designated reagents at the manufacturers recommended. Lipofectamine 3000 reagent thermo fisher scientific kr. Fugene hd transfection reagent for efficient mammalian cell.
Prepare plasmid dnalipid complexes recommend 2 doses of lipid. Vortex and incubate the mixture at room temperature for. Choosing a transfection reagent is easier when you can rely on the work of researchers who have used it before you. The day before transfection plate adherent cells in the amount that by the time of transfection. The protocols cover growing cells, culture conditions, and. Lfugene hd transfection reagent remains fully functional even.
This protocol takes around 2 months to complete, from mef isolation to ips cell establishment. Fugene hd transfection reagent quick protocol fb112pdf 99 kb english. These protocols were developed by promega corporation or fugent, l. Fugene 6 transfection reagent is a proprietary blend of lipids and other components supplied in 80% ethanol, sterilefiltered, and packaged in glass vials. The amount of dna and the fugene hd transfection reagent in protocol. Comparison of nanoparticlemediated transfection methods. Promega fugene hd transfection reagent fugene hd is a novel, nonliposomal reagent for transfecting dna into a variety of cell lines with high efficiency and low. Protocols fugene hd transfection reagent quick protocol fb112 pdf 99. Improving the success of your transfection promega connections.
Use the table below to find the right product for your experiment. Improving the success of your transfection promega. To transfect cells with sirna, follow the protocol as. Considering the transfection efficiency and lack of cytotoxicity in huvecs, it was established that. Different approaches and modifications such as the use of nanoparticles nps are being evaluated to increase transfection efficiencies. Several studies have focused on the attained transfection efficiency. The protocol does not require removal of serum or culture medium and does not require washing or changing of medium after introducing the reagentdna complex. Fugene 6, and fugene hd are trademarks of fugent, llc.
Fugene is a registered trademark of fugent, llc in the united states and other countries. Transitlt1 transfection reagent research reagent products. In addition to the protocol developed for the lipofectamine stem transfection reagent user guide weve created stem cellspecific protocols that will help guide you to high efficiency with low toxicity in individual stem cell types esc, nsc or msc and with your media of choice. Quick vortex, tap tube to settle, and sit 5 minutes at rt. Comparison of nanoparticlemediated transfection methods for. Since that time, its popularity has increased due to its ease of use, minimal to no cytotoxicity, and the high level of transfection in many different cell lines. Perform transfection into hek293t as above using fugene hd roche applied bioscience and harvest these cells 48 h later for use in. Always mix fugene 6 reagent prior to use vortex for one second or use inversion. Passaging and colony expansion of human pluripotent stem. Find msds or sds, a coa, data sheets and more information.
Quick transfection protocol preparing the fugene hd transfection reagent before use, allow fugene hd transfection reagent, dna, and diluent reach room temperature. For rapid analysis of your optimization results following this standard plate format, use the fugene hd transfection reagent optimization analysis worksheet. It is suitable for use in media with or without serum, and for transient or stable transfection, as well as cotransfections of multiple dna plasmids. Select the cell line, type of plate used and number of wells to be transfected and a protocol will be. Transfection efficiency and cytotoxicity of transfection reagents in. Plate cells so they will be 7090% confluent at the time of transfection. Fugene hd transfection reagent for efficient mammalian. It may also refer to other methods and cell types, although other terms are often preferred. For a list of conditions that have been used to transfect various cell types, visit the fugene hd transfection reagent protocol database. The two transfection reagents fugene 6 and fugene hd.
Helen sarantis, sergio grinstein, in methods in cell biology, 2012. For transient transfection, cells are typically assayed 2448 hours after transfection. This protocol is a slight modification of expression systems protocols for generating viruses. This is the protocol that i use when doing transfections with fugene 6. Fugene transfection protocol 1 add 3ul of fugene to 97ul of optimem, being sure to avoid touching the pipette tip to the side of the tube as the fugene will stick to plastic. Fugene hd protocol database the fugene hd protocol database is a collection of protocols that guide the transfection process for a variety of cell lines. Fugene 6 transfection reagent is stabilized for extended storage.
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