Transfection commonly refers to the introduction of nucleic acids into eukaryotic cells, or more specifically, into animal cells. Photograph of expected results dna lipofectamine scaling up or down. Life technologies corporation andor its affiliates warrant their. Application note gene delivery solutions stem cell. Genscript recommends using lipofectamine 2000 for all transfections.
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. Culture cells in serumfree media that are free of animal. Dmem invitrogen 11965092 with 10% fbs with no antibiotics 4. Descriptionlipofectamine 2000 is a proprietary formulation suitable for the transfection of. A hela cells were transfected with hcv dsrna using lipofectamine 2000 reagent. The intracellular trafficking mechanism of lipofectamine. Lipofectamine 2000 reagent is a lipidderived reagent designed to. The optimal level of each transfection reagent was determined empirically and all reagents were used according to manufacturers protocol. Invitrogen lipofectamine 3000 transfection reagent lipofectamine 3000 transfection reagent leverages our most advanced lipid nanoparticle technology to provide superior transfection performance with improved application outcomes and reproducible results.
Make dnalipofectamine 3000 complexes in serumfree medium such as. Lipofectamine 2000 transfection reagent from invitrogen. Make dna lipofectamine 3000 complexes in serum free medium such as. Lipofectamine rnaimax transfection reagent is a proprietary formulation for transfecting small rnas e. It consistently produces high transfection efficiency and high protein overexpression. Rna lipofectamine rnaimax complexes must be made in serum free medium such as optimem. Works effectively with all common cell lines as well as with many challenging ones, and can. Optimized conditions for successful transfection of human. Cd 293, 293 sfm ii, and vpsfm are media formulations known to inhibit. The lipofectamine 3000 is the bees knees though, i managed to get loads of transfects from a difficult to transfect cell line that had been giving us trouble with the lipofectamine 2000. What is the appropriate way of adding lipofectamine 3000 to cells. Test serumfree media for compatibility with lipofectamine 2000 since some. Lipofectamine 2000 transfection for hek293 cells about 6070% transfection rate transfect cells 6well plate.
Lipofectamine 2000 transfection reagent dartmouth college. Each reaction mix is sufficient for triplicate 96well, duplicate 24well, and single well 6well transfections, and accounts for pipetting variations. Invitrogen lipofectamine 2000 transfection reagent 1. 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. 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.
Product isguaranteed for 6 months from the date of shipment if stored properly. Lipofectamine 2000 reagent thermo fisher scientific. The animal origin free formulation ensures that mammalian cell culture and bioproduction processes are free of animalderived materials. Lipofectamine 2000 is a lipid based transfection reagent for eukaryotic cells. 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. 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. However, few studies have examined cellular responses to this delivery system. Refer to the cell lines database for a list of cell types successfully. Invitrogen lipofectamine 2000 transfection reagent. Lipofectamine or lipofectamine 2000 is a common transfection reagent, produced and sold by invitrogen, used in molecular and cellular biology. The microscopic analysis was performed using antirigi mab.
Lipofectamine reagents go online to view related products. Lipofectamine 2000 cd reagent is a proprietary animal originfree formulation for transfecting. Lipofectamine 2000 transfection reagent from thermo fisher. For lipofectamine plus transfections, the dna was preincubated with 4.
We tend to follow the protocol provided, which is simple and easy to follow. Test serumfree media formulations for compatibility with lipofectaminez 2000 as some serumfree formulations can inhibit cationic lipidmediated transfection. The instructions are recommending that you do this in duplicate with the two indicated different concentrations of lipofectamine 3000. Help frozen lipofectamine aug092006 hi i am really worried, i transported my lipofectamine from one lab to another in a dry ice container, and when i arrived to my lab and i open the box i found that the lipofectamine has been frozen i put it immediatly at 4 c, but i dont know if i damaged the lipofectamine can i still use this. When we add the fullserum media back 6 hours later, we use antibioticfree media. Lipofectamine 2000 dna transfection reagent protocol. I use 24well plates for transfections in triplicates. To determine the required amount of lipofectamine 2000 for forming of lipoplexes, different amounts of lipofectamine 2000, 1, 2, 4, 6. The purpose of this study is to evaluate the effect of sirna transfection using lipofectamine 2000 on cellular autophagy. Lipofectamine 2000 transfection reagent is versatile transfection reagent that has been shown to effectively transfect widest variety of adherent and suspension cell lines. Dnaplus lipid complexes must be made in serumfree medium such. Despite this, a satisfactory mechanismbased explanation of their. Optimizing transfection lipofectamine 2000 cd reagent.
Lipofectamine 2000 is commonly used for sirna transfections. Lipofectamine 2000 is then added to culture medium within a sterile tube along with the dna at a 3. I use lipofectamine 2000 invitrogen for transfection of both sirna and plasmid dna into these 293 cell lines. Lipofectamine 3000 protocol written out with timing. I tried reversetransfecting hepg2 cells using lipofectamine 2000. Transfections were performed in 24well plates using 0. Theres a difference between the two protocols lipofectamine 3000 has an extra reagent that needs to be added.
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. A lot of the transfection protocolefficiency depends on the kinds of cells you are using. Transient plasmid dna transfection into hela and hacat. Researchers use lipofectamine 2000 reagent for sirna and shrnabased gene knockdown experiments, as well as for gene expression studies. Lipofectamine or lipofectamine 2000 is a common transfection reagent, produced and sold by invitrogen. Dilute lipofectamine3000 reagent in optimem medium 2 tubes, mix well follow the instructions as indicated. Invitrogen lipofectamine 2000 transfection reagent is a proprietary formulation. Transfection of nih3t3 cells, hela, swis 3t3, 293t with lipofectamine 2000 1. For researchers interested in other delivery methods, we also offer additional invitrogen lipofectamine.
Plasmid dna and lipofectamine 2000 invitrogen, carlsbad, ca were diluted in two independent 250. Mix gently and incubate for 5 min at room temperature. Highest transfection efficiency in many cell types and formats e. Effects of lipofectamine 2000sirna complexes on autophagy. Transfection of nih3t3 cells, hela, swis 3t3, 293t with. Lipofectamine rnaimax has a broad peak of activity. Lipofectamine contains lipid subunits that can form liposomes in an aqueous environment, which entrap the. The trusted, simple, broadspectrum transfection reagent for most cell lines. Invitrogen lipofectamine 3000 transfection reagent. Although invitrogen recommends their own brand of culture medium for transfection, called optimem, we found no difference between this and conventional media. Lipofectamine 2000 from invitrogen biocompare product.
It is used to increase the transfection efficiency of rna including mrna and sirna or plasmid dna into in vitro cell cultures by lipofection. You can generally use other serumfree medium instead of optimem. Help frozen lipofectamine transfection and transduction. Transfect plasmid dna and sirna at the same time using lipofectamine 2000 reagent by adding 30 pmol 0. Invitrogen 11668019 view more versions of this product. Lipofectamine reagents are widely accepted as goldstandard for the safe delivery of exogenous dna or rna into cells. Versatile transfection reagent that has been shown to effectively transfect the widest variety of adherent and suspension cell lines. Lipofectamine 2000 and lipofectamine rnaimax transfection.1225 142 1006 788 970 707 1145 1050 1225 696 491 1438 105 331 1067 1332 572 832 1284 93 605 240 1110 1217 793 217 492