Lipofectamine ltx reagent is a plasmid transfection reagent that offers the right balance of potency and gentleness for your cells, resulting in the highest possible transfection efficiencies and viabilities. Lipofectamine 3000 reagent outperforms invitrogen lipofectamine 2000 and fugene hd reagents. The instructions are recommending that you do this in duplicate with the two indicated different concentrations of lipofectamine 3000. Theres the one with the blue cap, the one with the red cap, and the one with the green cap, and most people dont really understand the difference, but cell biologists seem to love these transfection reagents and are willing to pay a heavy premium for them. Alternatives to the megabrands of molecular biology tebu. Our lipofectamine reagentspecific protocols have been optimized for efficiency, viability, and reproducibility across a broad range of cell types see download.
Invitrogen lipofectamine rnaimax transfection reagent. Lipofectamine rnaimax transfection reagent is a proprietary formulation for transfecting small rnas \e. Following on from other products in the lipofectamine range, lipofectamine ltx is a lipid based reagent for transfecting dna and vector based rnai into eukaryotic cells. Follow manufacturers handbook protocol unless otherwise indicated. Invitrogen lipofectamine 3000 transfection reagent. Lipofectamine messengermax mrna transfection reagent delivers amazing transfection efficiency in neurons and a broadspectrum of primary cells, enabling improved application outcomes and more biologically. Lipofectamine transfection reagents offer efficient delivery of dna, rna. Lipofectaminez 2000 for transfection provides the following advantages.
A new transfection reagent for effective transfection of primary cells, hardtotransfect cells and sensitive established cell lines article pdf available april. Lipofectamine 2000 transfection reagent is a proprietary formulation for the transfection of nucleic acids dna and rna into eukaryotic cells and provides the following advantages. It is used to increase the transfection efficiency of rna including mrna and. Plate cells so they will be 7090% confluent at the time of transfection. Lipofectamine 3000 reagent is suitable for novel genome editing applications. Effect of cell density on total reporter gene activity. Invitrogen, usa were used for transfection of cell. It is used to increase the transfection efficiency of rna including mrna and sirna or plasmid dna into in vitro cell cultures by lipofection. Pdf optimization of sw480 colon cancer cells transfection with. Dilute lipofectamine3000 reagent in optimem medium 2 tubes, mix well follow the instructions as indicated. However, for cotransfection of sirnachimera and plasmid dna, lipofectamine 2000 should be used.
Use a range of several ul of lipofectamine and ug dna to increase viability after transfection and efficiency. Transfection using effectene, lipofectamine, and polyfect reagents many reagents are now available for optimization of cell transfection efficiency. You can search for similar products in our product directory. The lenti293t cell line has been tested as a suitable cell line for lentiviral production due to high transfection efficiency, high level of protein yield and high titer of lentiviral production. As a matter of fact, they should be evaluated in their entirety for the development of optimized nonviral gene delivery vectors. Alternatives to the megabrands of molecular biology. A h9 escs or b ipscs were transfected using lipofectamine 3000 reagent. Mix optimem with dna in an eppendorf 510 g dna0,8 ml optimem 2. Superior transfection efficiency requiring lower rnai concentrations, leading to more effective gene knockdown with minimal nonspecific effects. Many primary cell types have been successfully transfected with lipofectamine ltx reagent, including primary neural progenitor cells.
I use lipofectamine 2000 invitrogen for transfection of both sirna and plasmid dna into these 293 cell lines. The lenti293t is an hek 293 derived cell line stably expresses the sv40 large t antigen and neomycin resistance gene. Make sufficient 10% fbs in optimem media wo antibiotics sterile filter 2. Lipofectamine 3000 provided gentle but efficient transfection in several line of interest to our lab, heioc1 mouse cochlear 69fold, nccit and ntera2 3 fold testicular cancer lines and allowed for very nice dose response assays for cisplatin sensitvity assays. Invitrogen lipofectamine ltx reagent with plus reagent. The intracellular trafficking mechanism of lipofectamine. Lipofectamine 2000 reagent 11668027, invitrogen, lipofectin reagent. Effectene transfection reagent handbook qiagen lipofectamine transfection reagent handbook invitrogen polyfect transfection reagent handbook qiagen. The highest transfection efficiency in many cell types and formats e. Transfection protocol of lipofectamine rnaimax can be obtained from the invitrogen website.
The ratio of transfection reagent to episomal vectors was varied, and the. Highest transfection efficiency in many cell types and formats e. Prepare 6 well plates by adding bakedsterile 22x22 1. Lipofectamine 2000 dna transfection reagent protocol see page 2 to view a typical dna transfection procedure.
The intracellular trafficking mechanism of lipofectaminebased. Lipofectamine 3000 reagentefficient, reproducible transfection for biologically relevant cell models introduction with the increase of research in more biologically relevant cell models, existing molecular and cellular techniques need to be improved. Since launching in 1993, invitrogen lipofectamine reagents have become the most. You can generally use other serumfree medium instead of optimem.
Invitrogen lipofectamine 2000 transfection reagent life. Lipofectamine ltx reagent, when combined with plus reagent, provides the highest level of transfection in primary cells of any lipid or polymerbased reagent. Optimemin ul, equal volume for dna and lipofectamine lipofectaminein ul 25 protocol 6 well dish plate cells and grow to 70% confluence in 2 ml media without antibiotics dmem with 10% fbs, no psg. The following cell lines have been successfully transfected with the indicated transfection reagents. In tube b place 3 l lipofectamine ltx in 47 l of optimemi. Plate cells so they will be 6080% confluent at the time of transfection. Lipofectamine or lipofectamine 2000 is a common transfection reagent, produced and sold by invitrogen, used in molecular and cellular biology. Enhanced transfection efficiency using lipofectamine 3000 reagent. The transfection efficacy of lipofectamine 2000 was compromised by. Invitrogen lipofectamine 2000 transfection reagent. Sep 25, 2007 invitrogen have added a new product, lipofectamine ltx, to their extensive range of transfection products.
Lipofectamine crisprmax is more robust than lipofectamine rnaimax and several other transfection reagents in delivery of cas9 proteingrna complexes into a variety of cell lines. Lipofectamine ltx reagent is an advanced solution for gene expression in both common and challenging cell types. Comparative nucleic acid transfection efficacy in primary. Lipofectaminea rnaimax reagent thermo fisher scientific. Improved delivery of cas9 proteingrna complexes using. Lipofectamine contains lipid subunits that can form liposomes in an aqueous environment, which entrap the.
Lipofectamine is bad for cells we all know this but im obtaining % viability well over 100%. Pdf primary cells and certain established cells have generally been refractory to. Lipofectamine 3000 transfection reagent lipofectamine 3000 reagent leverages our most advanced lipid nanoparticle technology to enable superior transfection performance and reproducible results. Lipofectamine 3000 reagent maintains a high transfection efficiency within a robust dynamic range of lipid doses for quick and easy optimization. However, delivery of the cas9 rnps by neon electroporation was effective across all different cell lines tested and often was the only method that could produce indels. Given the vast selection of transfection reagents available today, the best option is usually to contact a technical specialist at tebubio for example for advice on the most suitable ones for your research. May 11, 2016 lipofectamine reagents are widely accepted as goldstandard for the safe delivery of exogenous dna or rna into cells. Lipofectamine 2000 transfection reagent is versatile transfection reagent that has been shown to effectively transfect widest variety of adherent and suspension cell lines. Lipofectamine messengermax mrna transfection reagent delivers amazing transfection efficiency in neurons and a broadspectrum of primary cells, enabling. Note, can adjust the amount of lipofectamine ltx and dna. Lipofectamine 2000 transfection reagent dartmouth college. Lipofectamine 2000 reagent thermo fisher scientific. Feb 18, 2016 lipofectamine crisprmax is more robust than lipofectamine rnaimax and several other transfection reagents in delivery of cas9 proteingrna complexes into a variety of cell lines.
Lipofectamine rnaimax reagent thermo fisher scientific. This work was aimed to understand better the mechanism of this phenomenon and its potential. Lipofectamine reagents offer you the ability to consistently transfect many cell types with high efficiency and exceptional ease of use. Optimemin ul, equal for dna and lipofectamine lipofectamine in ul 25. Use number of cells shown below, reduce media volume to 500 lwell, in tube a place 500 ng of dna in optimemi to a final volume of 50 l. Invitrogen lipofectamine 2000 transfection reagent 1. Invitrogen have added a new product, lipofectamine ltx, to their extensive range of transfection products. Enable high transfection efficiency in novel genome editing applications. Our lipofectamine reagentspecific protocols have been optimized for efficiency, viability, and reproducibility across a broad range of cell types see download protocol links in the table below. What is the invitrogen lipofectamine messengermax transfection reagent.
Lipofectamine ltx transfection reagent from invitrogen. Researchers use lipofectamine 2000 reagent for sirna and shrnabased gene knockdown experiments, as well as for gene expression studies. Effi ciency and ofp expression were analyzed 72 hours posttransfection and a u2os and b hepg2 cells showed 4fold and 80fold improvement with lipofectamine. Hiperfect qiagen, interferin polyplus, lipofectamine rnaimax invitrogen, lipofectamine 2000 invitrogen, dharmafect 1, dharmafect 2, dharmafect 3 and dharmafect 4 dharmacon, siport amine ambion, xtremegene roche and nter sigmaaldrich were used according to the. Lipofectamine 2000 is then added to culture medium within a sterile tube along with the dna at a 3. Upon optimization of transfection conditions, we observed 85, 75 or 55.
A new transfection reagent for effective transfection of. Lipofectamine is bad for cells we all know this but im obtaining %. For additional information on scaling your transfection reaction, see. Lipofectamine plasmid transfection bridges lab protocols. Advanced transfection with lipofectamine 2000 reagent. Invitrogen lipofectamine 3000 transfection reagent from. Lipofectamine o23, ot, hela, 2a1 wt shsy5y use 10x amount of dna polyfect 292t, hela, mcf7, ot70, otbag references. For additional information on scaling your transfection reaction, see page 4. Versatile transfection reagent that has been shown to effectively transfect the widest variety of adherent and suspension cell lines. In addition, invitrogen claims that lipofectamine 2000 gives the most proven.
During functional studies on the rat stressinducible hspa1b hsp70. Different cell lines were transfected with a gfp reporter vector using the indicated transfection reagents. Cells were stained for pluripotency with the ssea4 antibody, visualized by. Invitrogen lipofectamine 3000 transfection reagent 0. Vero cells respectively, while maximum achieved level of. Lipofectamine transient transfection of cos7 cells to. Adjust the amounts of components according to your tissue culture format. Lipofectamine contains lipid subunits that can form.
Lipofectamine rnaimax is proprietary animal originfree, chemically defined rnaispecific cationic lipid formulation designed specifically for delivery of sirna and mirna into all cell types. Lipofectamine 3000 protocol written out with timing. Lipofectamine transfection of c2c12 cells bridges lab. I use 24well plates for transfections in triplicates. Comparison of small interfering rna sirna delivery into. Lipofectamine 2000 lipofectamine 3000 competitor 1 competitor 2 western blot, hepg2 gststat. Dilute lipofectamine rnaimax reagent in optimem medium 2 dilute sirna in optimem medium 3 add diluted sirna to diluted lipofectamine rnaimax reagent 1. Delivery of episomal vectors into primary cells by means of. Protocol 6 well dish plate cells the day before so that they are at 9095% confluence in 2 ml media without antibiotics dmem with 10% fbs, no psg. Mix optimem and lipofectamine in another eppendorf 32 l lipofectamine in 0,8 ml optimem 3. Refer to the cell lines database for a list of cell types successfully transfected. Select the part name and then you can download the datasheet in pdf.
Lipofectamine 3000 reagent thermo fisher scientific sg. Laboratory 2010 lipofectamine 2000 transfection protocol 10cm plates reagents. Lipofectamine 2000 from invitrogen biocompare product. Despite this, a satisfactory mechanismbased explanation of their. Although invitrogen recommends their own brand of culture medium for transfection, called optimem, we found no difference between this and conventional media. Refer to the transfection collection and the invitrogen transfection guide available at for a list of cell lines and cell types that have been successfully transfected. Each reagent was used to transfect hek 293, hela, lncap, hepg2, and a549 cell lines in a 96well format, and gfp expression was analyzed 48 hours posttransfection. It increases the likelihood of successful cleavage and recombination with talens or crisprs through highly efficient transfection, ultimately maximizing the efficiency of genetic modifications and simplifying the downstream processes. Invitrogens lipofectamine 2000 is highly cited in the literature, a sign of a successful reagent, so when our lab embarked upon the path to develop a successful transfection protocol, we included lipofectamine 2000 along with the other leading reagents. Introducing the versatile reagent with the power to. Lipofectamine dna transfection reagents protocol see page 2 to view a typical plasmid transfection using a lipofectamine reagent. Optimemin ul, equal for dna and lipofectamine lipofectaminein ul 25. Invitrogen 11668019 view more versions of this product.