Applications

Transfection-EZ Biosystems: Broad Range Transfection Reagents

We offer a range of EZ Biosystems' general purpose transfection reagents for plasmid, siRNA, co-transfections, in vivo work and CRISPR. These reagents are cost-effective and have been specially formulated for maximum transfection efficiency of a wide range of different cell types with minimal toxicity.

Why broad spectrum for all type of nucleic acids?

Avalanche® transfection reagents have the broad spectrum for the transfection of large plasmid, mRNA, siRNA, and/or other type of nucleic acids. This feature makes Avalanche® transfection reagents the best option for co-transfection of different type and/or size of nucleic acids. The following are the reasons for the broad spectrum:

The key components of Avalanche® transfection reagents are mixtures of cationic lipids and cationic polymers, which contain all three types of amines: primary, secondary, and tertiary amines. The three types of amines have different positive charge strength, hence have different capabilities to combine with negative charged anions.

As anions, different types and sizes of nucleic acids have different densities and quantities of negative charges.

With optimized ratios of different cationic lipids and polymers, hence optimized ratios of the three different type of amines, together with optimized protocols, our Avalanche® mixtures have the optimized strength of positive charges that ensure them to efficiently combine with different type of nucleic acids of a variety of negative charge densities and quantities, and compact them into the right sizes of nanoparticles with the right densities of positive charge (zeta potential) on their surface. The optimized size and the right strength of zeta potential of the nanoparticles are the keys for the nanoparticles to interact efficiently with negatively charged cell surface, and trigger endocytosis process.

K2® Transfection System

DNA & RNA transfection kit for mammalian cells. Suitable for transfection of DNA, mRNA, siRNA und miRNA Suitable for cotransfection of DNA and RNA Especially suitable for human cells In many cases significantly increased transfection efficiency compared to common reagents Decreases the cells´ability to detect nucleic acids

Biontex

Catalogue No.	Description	Pack Size	Price	Qty	Change to: €
T060-0.2	K2® Transfection System	200 ul	POA	Quantity	Add to Order
T060-1.0	K2® Transfection System	1.5ml	£364.00	Quantity	Add to Order
T060-2.0	K2® Transfection System	2x1.5ml	£684.00	Quantity	Add to Order
T060-5.0	K2® Transfection System	5x1.5ml	£1,605.00	Quantity	Add to Order

Description

Highlights:

Suitable for transfection of DNA, mRNA, siRNA und miRNA
Suitable for cotransfection of DNA and RNA
Especially suitable for human cells
In many cases significantly increased transfection efficiency compared to common reagents
Decreases the cells´ability to detect nucleic acids

Eukaryotic cells have an innate immune system which makes them able to detect foreign substances such as lipopolysaccharides, bacterial or viral nucleic acids and proteins, and take action to inhibit invasion by potential pathogens. Moreover, the cells announce the presence of harmful substances to neighbouring cells via messenger molecules. As a consequence, these neighbours adopt defence modes without even having contact with the pathogens.

The finding that lipofection is also attacked by the defence measures of the innate immune system led us to develop the K2® Transfection System, consisting of the K2® Transfection Reagent and the K2® Multiplier. The K2® Transfection Reagent is based on powerful cationic lipids, while the Multiplier decreases the cells' ability to detect nucleic acids.

We were thus able to significantly increase transfection efficiency in many cases in comparison to traditional lipofection reagents, namely in transfection of DNA and mRNA. The K2® Transfection System is therefore suitable for transient and stable transfection of mammalian cell lines and primary cells with DNA (plasmids, bacmide), RNA (mRNA, miRNA, and siRNA), and modified nucleic acids (e.g. antisense oligonucleotides). Areas of application are particularly the production of proteins, antibodies, and viruses (e.g. adenoviruses, AAV, lentiviruses), cotransfections

of different nucleic acids, genome editing (e.g. CRISPR/Cas9, CRISPR/Cpf1) and gene silencing (gene knockdown).

COMPARATIVE STUDIES

DNA TRANSFECTION

Transfection efficiency of K2® Transfection System with pCMV-Luc compared with a commercial transfection reagent (L2K) and METAFECTENE® PRO under optimized conditions:

Transfection efficiency of K2® Transfection System with pCMV-Luc compared with L2k and METAFECTENE® PRO under optimized conditions

Transfection efficiency of K2® Transfection System with pCMV-GFP compared with a commercial transfection reagent (L2k) under optimized conditions:

Microscopy after transfection of A549 cells with K2® Transfection System and L2k

Microscopy after transfection of HeLa cells with K2® Transfection System and L2k

Microscopy after transfection of HT 1080 cells with K2® Transfection System and L2k

mRNA TRANSFECTION

Transfection efficiency of K2® Transfection System with Luciferase encoding mRNA compared with METAFECTENE® PRO under optimized conditions:

Transfection efficiency of K2® Transfection System with Luciferase encoding mRNA compared with METAFECTENE® PRO under optimized conditions

Transfection efficiency of K2® Transfection System with GFP encoding mRNA compared with METAFECTENE® PRO under optimized condition:

Microscopy after transfection of A549 cells with GFP encoding mRNA and K2® Transfection System compared to METAFECTENE® PRO

Microscopy after transfection of HeLa cells with GFP encoding mRNA and K2® Transfection System compared to METAFECTENE® PRO

Microscopy after transfection of HepG2 cells with GFP encoding mRNA and K2® Transfection System compared to METAFECTENE® PRO