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Cell Transfection:
Lipofectamine™ vs Transfectamine™


Transfection is the process of deliberately introducing naked or purified nucleic acids into eukaryotic cells by non-viral methods. It can be achieved by physical or chemical techniques.

Physical methods like electroporation or microinjection tend to have low efficiency and damage cells. Chemical-based transfection uses materials ranging from inexpensive calcium phosphate and organic dendrimers to advanced ones such as lipofection (lipid transfection) reagents like Lipofectamine™ and Transfectamine™, but must be optimized.

There are 2 key issues with transfection: one is to improve the transfection efficiency and the other is to minimize the cytotoxicity associated with performing the transfection process. Through rigorous in-house testing, we have successfully shown that cells transfected with Transfectamine™ 5000 provides much higher transfection efficiency with low cytotoxicity, as compared with similar reagents.

Transfection efficiency comparison in CHO-K1 cells. CHO-K1 cells were cultured in 6-well plate to ~90% confluency. 2.5 ug of GFP plasmid was transfected with Lipofectamine 2000, Lipofectamine 3000 and Transfectamine™ 5000. Images were taken 24 hours post transfection with fluorescent microscope through FITC channel.Transfection efficiency comparison in CHO-K1 cells. CHO-K1 cells were cultured in 6-well plate to ~90% confluency. 2.5 ug of GFP plasmid was transfected with Lipofectamine™ 2000, Lipofectamine™ 3000 and Transfectamine™ 5000. Images were taken 24 hours post transfection with fluorescent microscope through FITC channel.

 

 

Featured Products & Resources


AAT Bioquest provides a wide range of resources and specialized products for researchers transfecting cells in the lab, from Application Notes to FAQs.

AssayWise Letters Research Article:
Transfectamine™ 5000: An Efficient and Reliable DNA Delivery System

Drug Discovery Targets:
G-Protein-Coupled Receptors (GPCR)

Open-Access Tools
Online Calculators for DNA Research:
DNA Concentration Calculator
DNA Molecular Weight Calculator

GPCR activation of G protein coupled receptor V2R construct transfected cells with Vasopressin using FlexStation 3. The promiscuous G-protein Ga16 and G protein coupled receptor V2R construct were co-transfected in CHO-K1 cells following each transfection reagent's protocol. Intracellular calcium flux assay were performed ~36 hours post transfection. Cells were treated with vasopressin to induce rapid calcium flux and were observed using the calcium sensitive fluorescent dye Calbryte™ 520 AM.Activation of GPCR V2R construct transfected cells with Vasopressin. The promiscuous G-protein Ga16 and GPCR V2R construct were co-transfected in CHO-K1 cells by Lipofectamine™ 2000, Lipofectamine™ 3000, or Transfectamine™ 5000. Cells were treated with vasopressin to induce rapid calcium flux and were observed using the calcium sensitive fluorescent dye Calbryte™ 520 AM. Intracellular calcium flux assays were performed ~36 hours post-transfection.



 

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