We have had success using lipofectamine reagents in the past and the lipofectamine 3000 appears to be the best option. Use plus reagent to enhance transfection results with lipofectamine ltx reagent, lipofectamine reagent, lipofectin reagent, cellfectin reagent, and oligofectamine reagent. Each reaction mix is sufficient for triplicate 96well, duplicate 24well, and single well 6well transfections, and accounts for pipetting variations. These cyclooctyl compounds satisfy the above mentioned two. Designed for dramatically improved transfection efficiency, invitrogen lipofectamine stem transfection reagent offers a simple, robust and reproducible method for delivering dna, rna and rnp such as cas9grna complexes into a wide range of stem cells, including pluripotent stem cells, neural stem cells and mesenchymal stem cells figure 2. Pdf improved delivery of cas9 proteingrna complexes. Transfection protocols thermo fisher scientific in. L transfection of sirna to transfect cells with sirna, follow the protocol as described for dna but do not add p3000 reagent when diluting the sirna step 3. Lipofectamine 2000 reagent thermo fisher scientific. Invitrogen lipofectamine rnaimax transfection reagent. Reagent free and portable detection of bacillus anthracis spores using a microfluidic incubator and smartphone microscope. Transfection reagents, protocols and technical resources. Shop online for a wide selection of invitrogen lipofectamine rnaimax transfection reagent provides highest transfection efficiencies on widest variety of cell types for.
Transfection reagents are designed to deliver dna, small rnas, and proteins into eukaryotic cells. Estimation of disulfide bonds using ellmans reagent. 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 important guidelines. Nonlipid based chemical transfection reagent optimized for maximum transfection efficiency, easeofuse, and minimal cytotoxicity on a wide variety of mammallian cells. The intracellular trafficking mechanism of lipofectamine. Plate cells so they will be 7090% confluent at the time of transfection. Download fulltext pdf improved delivery of cas9 proteingrna complexes using lipofectamine crisprmax article pdf available in biotechnology letters 386 february 2016 with 624 reads. Lipofectamine rnaimax reagent thermo fisher scientific. Dna lipofectamine complexes must be made in serum free medium such as optimem reduced. This is often the best place to start, especially in a new cell line. Lipofectamine reagent is a proprietary formulation for transfecting nucleic acids into a wide range of eukaryotic cells.
Invitrogen lipofectamine 3000 transfection reagent. Transfection and genome engineering fisher scientific. Lipofectamine 3000 transfection reagent leverages our most advanced lipid nanoparticle technology to provide superior transfection performance with improved application outcomes and reproducible results. Lipofectaminea rnaimax reagent thermo fisher scientific. Make dna lipofectamine 3000 complexes in serum free. Transpass p transfection reagent should be stored at 4. Versatileone reagent for dna, rna, and cotransfection. May 11, 2016 the intracellular trafficking mechanism of lipofectaminebased transfection reagents and its implication for gene delivery. A high efficiency transfection protocol employing a common polycationic lipid is described. The procedure is based on the reaction of the thiol with dtnb to. Compound classes included phenols, thiols, vitamins, amino acids, proteins, nucleotide bases, unsaturated.
Lipofectamine 3000 reagent thermo fisher scientific kr. In order to evaluate transfection reagents under optimal conditions, test each reagent sidebyside on the same day with the same batch of cells. One certificate handles all products above, download pdf file. For common cell types, lipofectamine 3000 reagent is more costeffective than other reagents due to its high concentration and low usage amount. Transfection reagents in cloning and expression sigmaaldrich. A hela cells were transfected with hcv dsrna using lipofectamine 2000 reagent. Using lipofectamine, a widely used transfection reagent, we transfected 293t cells with a plasmid harboring the. Rna lipofectamine rnaimax complexes must be made in serum free medium such as optimem. This study shows that lipofectamine 2000 is an efficient reagent for the delivery of genes into sw480 cells. The amount of lipofectamine 2000 reagent required for successful transfection varies depending on the cell type and passage number. If you are unable to find a building block, reagent or any other product for your peptide synthesis projects, we welcome your request and will use it to broaden our product range even further.
Lipofectamine reagents are widely accepted as goldstandard for the safe delivery of exogenous dna or rna into cells. Polybrene infection transfection reagent tr1003 general guidelines. Pdf reagentfree and portable detection of bacillus. Alternatively, assays using reporter genes including gfp and luciferease, which are faster. This reagent provides an advanced solution for gene expression in common cell lines, particularly chinese hampster ovary cho, and challenging cell types such. Lipofectamine 2000 transfection reagent dartmouth college. Please bother us with your suggestions at matthias. Discover the versatile reagent with the power to transfect your most difficult cells for just pennies per reaction the invitrogen lipofectamine 3000 transfection reagent leverages our most advanced lipid nanoparticle technology to enable superior transfection performance and reproducible results. Store at 4 25 c product description tri reagent is a complete and readytouse reagent for the isolation of total rna or the simultaneous isolation of rna, dna and proteins from samples of human, animal, plant, yeast, bacterial and viral origin. Lipofectamine messengermax transfection reagent 11 expifectamine 293 transfection kit 14 broadspectrum transfection reagent 15 lipofectamine 2000 transfection reagent 15 electroporation 16 neon transfection system 16 in vivo delivery 17 invivofectamine 2. Cell type and reagent selection are amongst the most critical decisions for ensuring successful transfection with high efficiency and low cytotoxicity. Use the indicated volume of dna and plus reagent with each of the four volumes of lipofectamine ltx. Start any new transfection by testing the recommended four concentrations of lipofectamine 2000 reagent to determine an optimum amount. The handbook of organic analytical reagents, 2nd edition, is an indispensable source book of physicochemical.
Compare transfection reagents sidebyside using the same batch of cells. All chemfiles brochures can be found as freetodownload pdf file at. I personally have used this reagent to transfect cells for fluorescent microscopy, gst pull downs, and coimmunoprecipitation experiments. Not recommended for use with lipofectamine 2000 or lipofectamine rnaimax. If you find this doesnt work for your specific cell type, then you can you look to our cell specific protocols for further optimization. High efficiency gene transfer by multiple transfection protocol. Despite this, a satisfactory mechanismbased explanation of their. 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. This is necessary because serum is replete with rnases that degrade ribozymes. Lipofectamine ltx reagent is a plasmid transfection reagent that offers a balance of potency and gentleness for your cells, resulting in high transfection efficiencies and viabilities. Our lipofectamine reagent specific protocols have been optimized for efficiency, viability, and reproducibility across a broad range of cell types see download protocol links in the table below. Lipofectamine ltx reagent is advanced solution for gene expression in both common and challenging cell types. Download instruction manual hanna instruments singapore.
Animal free polycationic liposomal transfection reagent optimized for the transfection of hek293 cells. Lipofectamine reagents offer you the ability to consistently transfect many cell types with high efficiency and exceptional ease of use. It is used to increase the transfection efficiency of rna including mrna and sirna or plasmid dna into in vitro cell cultures by lipofection. Invitrogen lipofectamine ltx reagent with plus reagent 1ml. Jul 28, 2010 a thorough study was done to test the reactivity of the folinciocalteu fc reagent towards various compound classes. Our lipofectamine reagent protocols have been optimized for efficiency, viability, and reproducibility across a broad range of cell types. Lipofectamine 3000 reagent maintains a high transfection efficiency within a robust dynamic range of lipid doses for quick and easy optimization. Efficiently transfect difficulttotransfect cells lipofectamine 3000 reagent is designed to efficiently transfect difficulttotransfect cells, yielding superior transfection performance across the broadest array of cell types. 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. Invitrogen lipofectamine 3000 transfection reagent 1.
Prior to transfection, hek 293 cells are preincubated with serum free dmem medium for 1 h. The amount of protein and the incubation time with the transfection complex mixture used may be adjusted for optimal transfection efficiency. Use purified proteins and enzymes free of bacteria or toxins. Hek 293 cells are efficiently transfected with lipofectamine, with typically 70% of cells showing fluorescence following transfection with green fluorescent protein gfp. Optimization of the transfection condition generally, transfection optimization could be achieved by transfecting cells with sirnas targeting endogenous genes such as lamin ac and gapdh and then analyzing their expression by rtpcr or western blotting. Invitrogen lipofectamine 3000 transfection reagent from. Lipofectamine or lipofectamine 2000 is a common transfection reagent, produced and sold by invitrogen, used in molecular and cellular biology. Lipofectamine reagents go online to view related products. Fiesers reagents for organic synthesis pdf free download epdf. Compare transfection reagents transfection, electroporation. Lipofectamine 3000 reagent thermo fisher scientific tw. Rnalipofectamine rnaimax complexes must be made in serum free medium such as optimem. No other plasmid dnaspecific transfection reagent can match efficiency, convenience, and gentleness of lipofectamine ltx reagent for transfection of primary, challenging, and sensitive cell lines.
Lipofectamine stem transfection reagent thermo fisher. Bases are a very important class of reagents for synthetic organic chemistry, as in a large. A new transfection reagent for effective transfection of primary cells, hardtotransfect cells and sensitive established cell lines. Lipofectamine 2000 reagent is a proprietary formulation for transfecting nucleic acids into a wide range of eukaryotic cells. 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.
Dna lipofectamine ltx and plus complexes must be made in serum free medium such as optimem. Dnaplus lipid complexes must be made in serum free medium such. Gfp expression was analyzed 48 hours posttransfection. Whats the difference between lipofectine and lipofectamine. Invitrogen lipofectamine 3000 transfection reagent 0. Each reagent would need to be optimized before use i. Hi97734b cal check cuvette b for free and total chlorine hr. Blt erba liquid stable reagents instructions about the.
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