Invitrogen ltd inchinnan business park 3 fountain drive paisley pa4 9rf, uk tel. Invitrogen lipofectamine 3000 transfection reagent is the. The amount of lipofectamine 2000 reagent required for successful transfection varies depending on the cell type and passage number. Optimem i reduced serum medium lipofectamine rnaimax transfection reagent thermo fisher scientific 778150. In vitro and in vivo inhibition of rabies virus replication by rna.
Figure 1 expression results achieved with lipofectamine 2000 versus other. Invitrogen lipofectamine 2000 cd transfection reagent. Invitrogen lipofectamine 2000 transfection reagent is a proprietary formulation for the transfection of nucleic acids dna and rna into eukaryotic cells providing the following advantages. Lipofectamine stem reagent is compatible with a variety of media systems, including feeder free, helping support and simplify your stem cell culture workflow. An improved method for increasing the efficiency of gene. Lipofectamine 2000 reagent and lipofectamine ltx reagent. After a 5 minute incubation in optimem, the lipofectamine 2000 and dna are combined and incubated for 20 minutes with gentle shaking at room temperature.
Animal origin free formulation ensures that mammalian cell culture and bioproduction processes are free of animalderived materials. Comparison of k2 transfection system with lipofectamine. Dna lipofectamine ltx and plus complexes must be made in serum free medium such as optimem. Sep, 2005 both lipofectamine 2000 and dna are mixed with optimem media and the quantity of each is dependant on the type of culture vessel i. Comparison of small interfering rna sirna delivery into. Optimizing transfection of primary human umbilical vein. Us5627159a enhancement of lipid cationic transfections. Lipofectamine 2000 and lipofectamine rnaimax transfection. Dmem may be used to dilute lipofectaminez 2000, but transfection efficiency may be compromised. Lipofectamine 2000 is then added to culture medium within a sterile tube along with the dna at a 3. The transfection efficacy of lipofectamine 2000 was compromised by.
Researchers use lipofectamine 2000 reagent for sirna and shrnabased gene knockdown experiments, as well as for gene expression studies. Proprietary animal origin free, chemically defined formulation for transfecting nucleic acids into eukaryotic cells. Transfection of cultured eukaryotic cells using cationic lipid. Use a range of several ul of lipofectamine and ug dna to increase viability after transfection and efficiency. Procedural guidelines culture cells in serum free media that are free of animalderived components. Invitrogen lipofectamine 2000 transfection reagent is the mostcited lipidbased reagent in scienti. Highest transfection efficiency in many cell types and formats e. Jun 01, 2004 prior to mixing with dna, lipofectamine 2000 should be diluted in optimem i. Depending on the size of culture vessel being used, the lipofectamine 2000 is diluted between 25 and 50fold. May 11, 2016 briefly, complexes were prepared in optimem invitrogen, carlsbad, ca by mixing for each well of 6well plates 2. The trusted, simple, broadspectrum transfection reagent for most cell lines.
Lipofectamine 2000 cd reagent thermo fisher scientific. Lipofectamine 2000 from invitrogen biocompare product. Transfection commonly refers to the introduction of nucleic acids into eukaryotic cells, or more specifically, into animal cells. Lipofectamine 2000 transfection reagent life technologies, cat. Lipofectamine 2000 dna transfection reagent protocol. Jpy 1,9202,750 neon invitrogen tips special buffers cost per sample. The solutions were combined and mixed by pipetting up and down once. Lipofectamine or lipofectamine 2000 is a common transfection reagent, produced and sold by invitrogen, used in molecular and cellular biology.
Forward transfection use this procedure to forward transfect sirna into mammalian cells in a 24well format for other formats, see scaling up or down transfections, page 3. Lipofectamine 3000 reagent outperforms lipofectamine 2000 and. Test serum free media formulations for compatibility with lipofectaminez 2000 as some serum free formulations can inhibit cationic lipidmediated transfection. It is very easy to use, somewhere between lipofectamine and fugene 6.
It is used to increase the transfection efficiency of rna including mrna and sirna or plasmid dna into in vitro cell cultures by lipofection. In columns 11 and 12, 100 l of lipofectamine 2000 and serumfree. Lipofectamine 2000 reagent thermo fisher scientific. Lipofectamine 2000 and lipofectamine ltx invitrogen, carlsbad, ca, usa. I use lipofectamine 2000 invitrogen for transfection of both sirna and plasmid dna into these 293 cell lines. Lipofectamine 3000 transfection reagent 10fold higher transfection ef. Lipofectamine 2000 lp2000, invitrogen is used as the transfection reagent, and the optimized condition for chimera rnai transfection should be determined. Allow the solution to spread over the plate and then remove it immediately via vacuum aspiration. A method is provided for enhancing transfection efficiency of eukaryotic cells comprising contacting said cells with a lipid aggregate comprising nucleic acid and a polycationic lipid composition in the presence of a polycationic compound and serum. Lipofectamine ltx and plus reagent are proprietary formulations for transfecting nucleic acids into a wide range of eukaryotic cells. A11803 lipofectamine 2000 transfection reagent life technologies, cat. Researchers can achieve up to 80% or better transfection efficiency in pluripotent stem cells pscs and neural stem cells nscs, and up to 45% in meschenchymal stem cells mscs. Test serum free media formulations for compatibility with lipofectamine 2000 as some serum free formulations can inhibit cationic lipidmediated transfection. Invitrogen lipofectamine 2000 transfection reagent 0.
Test serum free media formulations for compatibility with lipofectamine 2000 as some serum free formulations can. Photograph of expected results dna lipofectamine scaling up or down. Invitrogen lipofectamine 2000 transfection reagent is a proprietary formulation. The product manual includes advice on how to optimize transfection. After 30 minute incubation, add 100 l of the dna lipofectamine ltx reagent complexes directly to each. Advanced transfection with lipofectamine 2000 reagent.
Invitrogen lipofectamine 2000 transfection reagent is a versatile transfection reagent that has been shown to effectively transfect a variety of payloads into the widest variety of adherent. Lipofectamine 2000 reagent and lipofectamine 3000 reagent were used to transfect 17 cell lines with a gfpexpressing plasmid in a 24well plate format using 0. Test serum free media for compatibility with lipofectamine 2000 cd reagent since some serum free formulations e. Lipofectamine 3000 reagent is a proprietary formulation for transfecting nucleic acids into a wide range of eukaryotic cells and especially designed for hard to transfect cells. Select sirna designs will be provided free of charge.
Although invitrogen recommends their own brand of culture medium for transfection, called optimem, we found no difference between this and conventional media. Jpy 210 nucl eofect or lon za cuvettes special buffers cost per sample. For additional information, and protocols on transfecting suspension cells refer to the manual at. Us5627159a enhancement of lipid cationic transfections in. Lipofectamine reagents go online to view related products. Achieve 99% transfection efficiency in a single stroke. It is not necessary to remove complexes or changeadd medium after transfection, but complexes may be removed after 46 hours. Invitrogen 11668019 view more versions of this product. Start any new transfection by testing the recommended four concentrations of lipofectamine 2000 reagent to determine an optimum amount. Jul 17, 2007 the lipofectamine 2000 and the dna are diluted in separate polypropylene tubes with a small volume of serum free media.
Lipofectamine 3000 reagent maintains a high transfection efficiency within a robust dynamic range of lipid doses for quick and easy optimization. The plasmid was eluted in nucleasefree h2o promega. Mix optimem and lipofectamine in another eppendorf 32 l lipofectamine in 0,8 ml optimem 3. Pellet was airdried for 5 minutes and dissolved in. Lipofectamine rnaimax has a broad peak of activity. Dna solution, mix gently and incubate 30 minutes at room temperature to form dna lipofectamine ltx reagent complexes. The intracellular trafficking mechanism of lipofectamine. Cell culture hek293t cells were cultivated in dmem invitrogen supplemented with.
Dmem may be used to dilute lipofectamine 2000, but transfection efficiency may be compromised. Mix by pipetting and incubate for 5 to 10 min at room temperature. Dna lipofectamine 2000 cd complexes can be added directly to cells in culture medium. The chimera rnai is thawed make aliquots of chimera rnai 10. Lipofectamine rnaimax transfection reagent is a proprietary formulation for transfecting small rnas e. Gibco highquality selection agents provide unique solutions for your. Lipofectamine 2000 cd reagent is a proprietary animal originfree formulation for transfecting nucleic acids into eukaryotic cells. Lipofectamine 2000 cd reagent is a proprietary animal origin free formulation for transfecting nucleic acids into eukaryotic cells. Invitrogen lipofectamine 2000 transfection reagent.
Disposable kits special buffers nepa21 nucl eofect or neon. The animal origin free formulation ensures that mammalian cell culture and bioproduction processes are free of animalderived materials. I have used lf2000 successfully with the following cell lines. Remove culture medium and replace with serum free optimem medium in co2 incubator. Optimizing transfection lipofectamine 2000 cd reagent. The polycationic compound is preferably polybrene, and the lipid aggregate preferably comprises liposomes of nucleic acid and lipofectamine. I get consistent results using this lipid for my transfections. Lipofectamine 3000 reagent thermo fisher scientific. Sep 25, 2007 lipofectamine 2000 lf2000 is a reagent that i have found fits exactly what i need. Versatile transfection reagent that has been shown to effectively transfect the widest variety of adherent and suspension cell lines. Lipofectamine 2000 transfection reagent is a versatile transfection reagent that has been shown to effectively transfect the widest variety of adherent and suspension cell lines. Test serum free media for compatibility with lipofectamine 2000 since some. Rnaimax transfection reagent 778030, invitrogen, waltham, ma, usa.
Make dna lipofectamine 3000 complexes in serum free medium such as. Transfection of exogenous gene in specific plasmids to various cells is. I use 24well plates for transfections in triplicates. Dna lipofectamine complexes must be made in serum free medium such as optimem reduced serum medium and can be added directly to cells in culture medium, in the presence or absence of serumantibiotic. Lipofectamine contains lipid subunits that can form liposomes in an aqueous environment, which entrap the. Transfection efficiency in a cancer cell line panel. Gibco psc neural induction medium is a serum free medium that provides high efficiency neural induction of human pluripotent stems cells in only 7 days. Invitrogen transfection guide available at for a list of cell. Comparison of k2 transfection system with lipofectamine 2000. Rna lipofectamine rnaimax complexes must be made in serum free medium such as optimem. Lipofectamine 2000 transfection reagent from invitrogen.
So far, i have been very impressed by the performance of lipofectamine 2000. After a 5 minute incubation, the two volumes are mixed, and the tubes incubated at room temperature for 20 minutes to allow the reagentdna complexes to form. Lipofectamine 2000 cd reagent 12566014 1 ml 4c 1 do not freeze. Lipofectamine 2000 and lipofectamine rnaimax transfection reagents introduction life technologies is committed to providing researchers and scientists the best products, enabling cutting edge technology while keeping the environment in mind. Transfect plasmid dna and sirna at the same time using lipofectamine 2000 reagent by adding 30 pmol 0. Gently mix dna and lf2000, incubate at room temperature for 20 minutes to allow dna and lipid to form complexes.
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