Background Introduction

Transfection enables us to better understand how target genes are expressed and regulated in cells. Common transfection methods currently include calcium phosphate co-precipitation, electroporation, DEAE-dextran and polybrene, mechanical methods (such as microinjection and gene gun), and cationic liposome transfection reagents, among which cationic liposome transfection reagents are commonly used.

 

 

Figure 1: Schematic Diagram of Different Transfection Methods (Image source: Daily Biology Reviews)

 

Comparison of Different Transfection Methods 

Transfection Method	Advantages	Disadvantages
Calcium Phosphate Co-precipitation	Low cost, relatively simple operation	Unstable transfection efficiency, prone to issues such as DNA aggregation affecting transfection efficiency, high cytotoxicity, poor transfection efficiency
Electroporation	High efficiency, suitable for all types of cells	Expensive electroporation equipment, high cell death rate, requires large amounts of nucleic acids and cells
DEAE-Dextran	Effective for transfection of adherent cells, can also be used for certain suspension cells, simple operation, repeatable results	Cell preference, some toxicity to cells, serum inhibition of cell growth required during transfection
Polybrene	Relatively simple operation, moderate price	Low efficiency, some limitations in application
Cationic Lipid Reagents	Simple operation, high efficiency, wide applicability, good repeatability, low toxicity	Slightly higher price compared to other chemical reagents
Viral Transduction	High efficiency, effective for difficult-to-transfect cells, good transduction in primary cells	Complex viral packaging process, some risk associated with viruses
Polyethylenimine (PEI)	Low price, simple operation, wide applicability	Lower efficiency compared to lipid-based transfection reagents, some cells exhibit sensitive reactions after contact

Celthy Trans liposome nucleic acid transfection reagent, as a cationic lipid transfection reagent, has higher transfection efficiency and more convenient operation compared to the widely used Lipo2000. It has been validated in various cell lines, as summarized in the list of validated cell lines below.

 

Mechanism of Action

Cationic lipids can wrap nucleic acids through electrostatic interactions due to the positive charges on their surface, forming nucleic acid-liposome complexes. The cell membrane surface carries a negative charge, allowing the complex to be adsorbed. Once adsorbed, the complex can enter the cell through membrane fusion or endocytosis, forming inclusion bodies within the cell. A small portion of DNA can be released from the inclusion bodies and enter the cytoplasm, further entering the cell nucleus for transcription and expression.

 

 

Figure 2: Liposome-mediated Transfection and Endocytosis Process

 

Celthy Trans Transfection Reagent Product Features

High efficiency: capable of transfecting most eukaryotic cells, with transfection efficiency exceeding 90% for common cell lines;

Low toxicity: transfected cells exhibit good morphology and express high levels of transfected gene proteins;

Convenient operation: liposome complexes can be directly added to serum-containing culture media;

Versatility: suitable for both transient and stable transfection experiments.

 

Operation Procedure

Figure 3: Cell Transfection Experimental Workflow

 

Product Data

 

Figure 4: Dual Plasmid Transfection Reagent Validation Results

 

Summary of Validated Cell Lines

293T	293FT	HEK293	HEK 293T	Caco2
COS-7	DF-1	Lenti X-293T	HeLa	HepG2
LM3	MCF-7	MDA-MB-231	MEF	NCI-H1975
NIH-3T3	PC12	Raw264.7	Vero	H9c2
Hepa1-6	MCF10A	CHO	5-8F	BV-2
A549	3t3	H520	HUVEC	C6
HaCaT	N2A	SGC-7901	H9	MKN-28
Hep 3B	SMCC7721	RKO	C2C12	More…

 

Customer Feedback Results Presentation

Celthy Trans Liposomal Transfection Application

Activation of Cell Autophagosome Formation by Bacterial Protein Cell: HeLa Transfected Plasmid: Bacterial Effector Protein A LC3II: Autophagosome Marker Protein		Hoechst 33342 Spindle Structure during Cell Division Cell: HeLa Transfected Plasmid: Flag-tubulin Nuclear Staining: Hoechst 33342
Celthy Trans	Lipo3000	Celthy Trans	Lipo2000
Image Source: Fudan University Cell Line: HEK293 Nucleic Acid Type: DNA		Image Source: Changchun Institute of Applied Chemistry Cell Line: HeLa Nucleic Acid Type: DNA

Flow Cytometry Fluorescence Observation Results

Celthy Trans (13.27%)	Lipo2000 (8.64%)	FuGenHD (9.24%)
Cell Line: C2C12 Fluorescent Protein: EGFP (Enhanced Green Fluorescent Protein) Nucleic Acid Type: DNA Data Source: South China Agricultural University

 

Partial Cell Transfection Data Provided by the Customer (For Reference Only)

Cell	Culture Plate	Cell Density	DNA	Celthy Trans	Efficiency
293T	6 well	80%	1μg	2μL	90%
293FT	24 well	85%	1μg	4μL	90%
Hela	12 well	90%	0.2μg	0.6μL	90%
Raw264.7	35mm plate	80%	1μg	2μL	90%
Hek293	6 well	95%	2μg	10μL	80-90%
NCI-H1975	6 well	80%	4μg	10μL	80%
3t3	12 well	90%	1μg	5μL	50%

 

FAQ

Q1: Can serum be present when preparing the nucleic acid transfection reagent complex?

a: The presence of serum can affect the formation of liposomes. It is recommended to use serum-free medium (usually MEM medium) when preparing the nucleic acid transfection reagent complex.

Q2: Is it necessary to change to serum-free medium during the transfection experiment?

a: No, our transfection reagent can be used for transfection in media containing serum.

Q3: Is it necessary to terminate the transfection after transfection?

a: No. The liposome complex can remain stable for 6 hours. If the cell medium has not been changed before transfection, it is necessary to change to fresh medium after 4-6 hours to ensure the nutrients required for normal cell growth. However, if the medium has been changed before transfection, it is not necessary to change the medium again after liposome transfection.

Q4: Precautions for storing and using the transfection reagent?

a: This reagent must be stored at 4°C. Avoid repeated long-term opening of the bottle, as prolonged exposure to air can cause oxidation of the liposomes and affect transfection efficiency.

Q5: What should be noted to improve transfection efficiency?

a: Cell density during transfection, 90%-95%.

b: Use MEM serum-free medium for dilution of nucleic acids and liposome diluents during transfection.

c: Medium change can be performed 4-6 hours after transfection.

 

Related Products

Product Name	Catalog number	Specification
Celthy 2000 Liposomal Transfection Reagent	C130002S	0.5 mL
	C130002M	1.0 mL

 

Application Scenario	Product Name	Catalog number	Specification
Cell Type: Adherent/Suspension Nucleic Acid Type: DNA, siRNA	Celthy 2000 Liposomal Transfection Reagent	C130002S	0.5 mL
		C130002M	1.0 mL
Cell Type: Adherent/Suspension Nucleic Acid Type: miRNA, siRNA,DNA	Celthy 3000 Liposomal Transfection Reagent	C130003S	0.5 mL
		C130003M	1 mL