Transfecting HEK293T cells with H5N1 pseudotypes [HIV Core + luciferase].
Endofectin Max produced greater RLU values at a lower HA conc. comapred to PEI and the vice versa when using high HA conc.
Dear Shabaz,
Transfection methods:
There is a wide range of transfection methods that include physical, chemical and biological techniques. These techniques generally involve the use of transient or stable transfection methods to incorporate nucleic acids into cells. Transient transfection techniques involve the introduction of DNA into cells, but in this method, the DNA does not integrate with the cellular chromosomes. This technique facilitates high transfection efficiencies and the gene transcripts can be analyzed after a period of 1-4 days. For large-scale transient gene expression (TGE) in mammalian cell cultures, transfection vehicles such as polyethylenimine (PEI) and calcium phosphate (CaPi) can be used. Furthermore, large-scale TGE methods have also been developed using Chinese hamster ovary (CHO) cells in the absence of serum1. Stable transfection techniques involve the integration of the transfected DNA into cellular chromosomes or the formation of episomes. The stably transfected cell can be subsequently identified using selectable markers such as dihydrofolate reductase (DHFR), hygromycin B phosphotransferase (HPH) and adenosine deaminase (ADA) among several others. Some of the commonly used transfection techniques include calcium phosphate precipitation, lipofection, electroporation, and viral delivery. Additionally, these methods can be used in cotransfections. These techniques involve the simultaneous delivery of two distinct nucleic acids into the same cell and are often used to achieve stable transfections. Transfections methods have evolved to include several new methods such as the biolistic delivery systems that use high velocity microparticles to deliver nucleic acids in cell, and in vivo transfection protocols that facilitate systemic delivery of siRNA molecules.
EndoFectin™ Max is a lipid-based transfection reagent. This versatile transfection reagent has been tested and optimized for highly efficient transfection of a wide collection of commonly used cell lines as well as many difficult-to-transfect cell lines.
Features:
Superior transfection efficiency for a broad range of cell lines compared with commonly used transfection reagents
Low cytotoxicity
Does not require removal of serum or culture medium
Does not require washing or changing medium after transfection
For overexpression, knockdown, knockout, as well as high-throughput applications.
Comparison of the chemical struures of EndoFectin with polyethylenimine (PEI) reveals that the former is more lipophilic than the latter. This cgaracteristic makes the reagent more permeable to cells and thus better transfection is expected.
For more details on these reagents, please use the following links:
http://www.biocompare.com/11119-Chemicals-and-Reagents/7936635-EndoFectin-Max-Transfection-Reagent/?ncatid=11231&ppim=7936635_1_0
http://www.sigmaaldrich.com/technical-documents/articles/biology/transfection-reagents.html
hoping this will be helpful,
Rafik
Thank you Rafik that helped out a lot. I just wanted to ask if you have a reference regarding the lipophilic structure of endofectin max
Cheers,
Shabaz
The kinetics of PEI is also quite slow. You will have to wait longer to see maximal expression than with other agents
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