Increase transfection efficiency and cell viability with. Transfection efficiency was assessed by facs analysis in various cell lines 24 h after transfection in 24well plates. Lipofectamine 2000 cd reagent is a proprietary animal originfree formulation for transfecting nucleic acids into eukaryotic cells. A preoptimized a549 transfection kit is available from altogen biosystems, which includes. A549 chok1 cos7 hek 293 hela hepg2 keratinocytes lncap neuro2a nih 3t3 pc3 fe 3. Introducing the versatile reagent with the power to. Lipofectamine 2000 right panel around 5% efficiency.
The commonlyused plasmid dna or rna lipofectamine transfection reagent had been shown to be able to deliver many proteins into cells, so we tested several lipofectamine variants for their ability to efficiently deliver cas9 rnps sells et al. How to maximize transfection efficiency in a549 cells. Lipofectamine 3000 reagent maintains a high transfection efficiency within a robust dynamic range of lipid doses for quick and easy optimization. Transfection protocol of adherent a549 cells 96well plate. Transfecting plasmid dna into a549 cells using lipofectamine 3000 reagent. Each reagent was used to transfect hek 293, hela, lncap, a549, and. Viafect transfection reagent allows highefficiency transfection of a wide range of. Altogen biosystems provides in vivo transfection reagents, over 100 preoptimized in vitro transfection kits for cell lines and primary cells, and electroporation delivery products. Figure 3 illustrates differences in expression and toxicity profiles for two common cell types, a549 and mdck cells.
Choosing the right transfection reagent for optimal efficiency. A comparison showing transfection efficiency of polyjet reagent vs. I have used lipofectamine and fugene for plasmid transfection. Lipofection has become more common than the capo 4 precipitation, because it is just as simple to use but frequently provides a much better transfection efficiency. Cell transfection in a 24well plate using crisprmax. I use lipofectamine 2000 invitrogen for transfection of both sirna and plasmid dna into these 293 cell lines. Lipofectamine 2000 transfection reagent dartmouth college. A reverse transfection protocol was used to transfect mcf7 and hepg2 cells.
Transfection efficiencies were assessed by gfp signal. Comparative transfection efficiency of jetprime versus its main competitor. Achieve superior transfection efficiency while maintaining low toxicity lipofectamine 3000 reagent maintains a high transfection efficiency. Dna transfection for luciferase assay in human alevolar. Different cell lines were transfected with a gfp reporter vector using the indicated transfection. A549, carcinomic human alveolar basal epithelial cells.
Igur high efficiency transfection on a broad range. Enhanced transfection efficiency using lipofectamine 3000 reagent. Mirus lipofectamine 2000 and oligofectamine invitrogen. Transfection optimizationhow to get it right biocompare. I use 24well plates for transfections in triplicates. I found lipofectamine and geneporter2 might not only damage a549 also have worse tranfection efficiecy, so i used jetpei to transfection.
A new transfection reagent formulated specifically for a549 cells. Copy link enlarge image comparison of forward and reverse transfection in a549. It is no surprise that lipoplexes stick to the plastic surfaces of cell culture. To transfect cells with sirna, follow the protocol as described for dna but. Singlecell transfection by electroporation using an. On the day of transfection just prior to preparing complexes, plate 1. Transfection the delivery of dna or rna into eukaryotic cells is a powerful tool used to study and control gene expression. Rapid and highly efficient mammalian cell engineering via. Xfect provides excellent transfection for most cell types. Singlecell transfection of adherent cells has been accomplished using singlecell electroporation scep with a pulled capillary. One day prior to transfection, adherent cells were plated onto 24well plates at 0. Transfection guide overview of transfection methods. A549 dharmafect 2 hct116 dharmafect 1 huh7 dharmafect 3 a673 lipofectamine 2000. Transfection efficiencies lower than 50% were not reported in this table.
On the day of transfection, which should be 1 day following cell plating, perform the following steps, which have been optimized for a single well of a 24well plate using lipofectamine 3000 transfection reagent. Hela cells typically transfect with 8590% efficiency. A549 transfection kit lung carcinoma, ccl185 altogen. Transfection protocol lipofectamine 3000 transfection. I have been transfecting a549 cells for long time but still could not get transfection efficiency more than 30%. Lipofectamine 3000 and rnaimax outperformed lipofectamine 2000. Transitx2 dynamic delivery system and lipofectamine 2000 transfection reagent were used to. Viafect reagent provided optimal transfection efficiency and low toxicity for the majority of the cell lines tested, although there were some cell lines e. Hepesbuffered physiological saline solution containing. Transient plasmid dna transfection into hela and hacat. However, significant toxicity from lipofectamine 2000 was also reported in. Viafect reagent offers performance similar to or better than that of lipofectamine 2000 reagent, and it may be suitable.
Transfection protocol of adherent a549 cells 96well plate 1. This reagent delivers superior transfection efficiency. I have used 3 kinds of transfection reagents lipofectamine, geneporter2, jetpei to test which one was better for a549 cell. The reagent provides high transfection efficiency in many cells types and formats. Protocol for lipidmediated transient transfection in a549 epithelial lung cell line.
Improved delivery of cas9 proteingrna complexes using. 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. Transitx2 transfection reagent a dynamic transfection system for the. Ccl185 human lung carcinoma cells by fugene hd in 96 well plates. Endofectin transfection reagent, cell lines genecopoeia. The optimized protocol for a 24well plate to transfect a549 cells is described below. L growth medium for a single well of a 24well plate. Xfect is a biodegradable transfection polymer, screened from a pool of 2,300 candidates for its very high transfection efficiency and low cytotoxicity profile. Cloned genes can be transfected into cells for biochemical characterization. Lipofectamine 2000 transfection, the expression of gfp was detected in 76,4% of hela cells with a mean fluorescence intensity of 437. Cell plating a549 cells were seeded from 90100% confluent culture the day before the transfection. This reference provides a recommended procedure to transfect plasmid dna into human a549 lung carcinoma cells atcc, cat. Transfection guide overview of transfection methods promega. Transfection reagents are highly efficient for dna and sirna transfection.
Invitrogen lipofectamine 3000 transfection reagent. N v i s i on io 20 transit certified transfection reagents. A549 lipofectamine 3000 lipofectamine 2000 fugene hd lipofectamine 2000 lipofectamine 3000 competitor 1 competitor 2 western blot, hepg2 gststat. Protocol for lipidmediated transient transfection in a549 epithelial. A b on the day of transfection, which should be 1 day following cell plating, perform the following steps, which have been optimized for a single well of a 24well plate using invitrogen lipofectamine 3000 transfection. From expert transfection reagent developer molecular transfer inc. Transfecting plasmid dna into a549 cells using lipofectamine. In biontex experiments gfp positivity increased to 90. In hacat cells, lipofectamine 2000 achieved a transfection efficiency. Invitrogen lipofectamine 3000 transfection reagent 1. For additional information on scaling your transfection.
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