Celthy Liposome Transfection Reagent, Confidence in Transfection

Celthy Liposome Transfection Reagent, Confidence in Transfection

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


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