"Heyuan Liji" is a joint venture between Heyuan Biotechnology (stock code: 688238) and Liji Biotechnology, specializing in the "Liji Biotechnology" and "Life-ilab" reagent brands
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EZ Trans Lipo Cell Transfection Reagent (Liposome) (hereinafter referred to as "EZ Trans Lipo") is the latest cell transfection reagent developed by Li Ji Biology. EZ Trans Lipo wraps nucleic acid in nano liposomes and uses a special delivery mechanism to deliver exogenous DNA, RNA, and siRNA into various cells. It can be transfected into adherent cells, suspended cells, and can also be used for siRNA and shRNA gene knockout experiments and gene expression studies.
After comparing and testing nearly a hundred types of cells, EZ Trans Lipo showed the same or better transfection efficiency and cytotoxicity performance as imported products in the vast majority of the tested cells; It has the advantages of high transfection efficiency, low cytotoxicity, wide applicability to cells, and high cost-effectiveness.
Product Name | Specifications |
EZ Trans Lipo Cell Transfection Reagent (Liposome) | 1 mL |
Blue ice transportation. Stored at 4 ℃, with a shelf life of 12 months.
Transfection method | principle | Main applications | characteristic |
Cationic polymer | Positively charged polymers form positively charged complexes with negatively charged phosphate groups in nucleic acids, which then interact with negatively charged proteoglycans on the cell surface and enter the cell through endocytosis. | Stable transfection Instantaneous transfection All cells | In addition to the high transfection efficiency, simple operation, wide applicability, and good repeatability of cationic liposomes, it also has the characteristics of high transfection efficiency and low cytotoxicity in vivo, making it a new generation of transfection reagents. |
Cationic liposome method (EZ Trans lipo pro) | Positively charged liposomes form complexes with negatively charged phosphate groups in nucleic acids, and then the remaining nuclei on the liposomes bind to the negatively charged nuclei of sialic acid residues on the cell membrane; In addition, cells can also engulf DNA encapsulated nanoliposome particles into cells through endocytosis. | Stable transfection Instantaneous transfection All cells | The usage method is simple, it can carry large fragments of DNA, and is applicable to various types of naked DNA, RNA, or siRNA. It can be transfected into various types of cells without immunogenicity. It has high efficiency in in vitro gene transfection. |
DEAE glucan method | The complex formed by the interaction between positively charged DEAE glucan and negatively charged phosphate backbone of nucleic acid is internalized by cells | Instantaneous transfection | Relatively simple and reproducible compared to calcium phosphate, but it has certain toxic side effects on cells. Serum needs to be removed during transfection and is generally only used for BSC-1, CV-1, COS cell lines |
Calcium phosphate method | Calcium phosphate DNA complex adsorbed onto cell membrane and internalized by cell | Stable transfection Transfection with dye | Not suitable for primary cells (requiring high DNA concentration), easy to operate but with poor repeatability. Some cells are not suitable. CSCL gradient centrifugation is recommended for cells, as transfection requires a high copy number |
Retrovirus (RNA) | It enters the host cell through the interaction between the virus membrane glycoprotein and the receptor on the host cell surface, and then reverses into an enzyme to initiate the synthesis of DNA, which is randomly integrated into the host genome | Stable transfection Specific host cells | Can be used for difficult to transfect cells, primary cells, in vivo cells, etc., but the gene carrying capacity should not be too large |
adenovirus (Double stranded DNA) | First, it binds to the receptors on the cell surface, and then is engulfed by the cell under the mediation of α v integrin | Instantaneous transfection Specific host cells | Can be used for difficult to transfect cells, safety factors need to be considered |
Biolistic Particle Transfer Method (Gene gun particle bombardment method) | The DNA is precipitated using microscopic heavy metal particles, and then the coated particles are projected into the cell using a ballistic device. The DNA is gradually released inside the cell and expressed | Transient transfection Stable transfection | Can be used for: human epidermal cells, fibroblasts, lymphocyte lines, and primary cells |
Microinjection method | Directly inject DNA into the target cell nucleus through microscopic manipulation | Stable transfection Instantaneous transfection | The number of transfected cells is limited, and they are mostly used for engineering or transgenic animal embryonic cells |
Electroporation method | High pulse voltage disrupts the cell membrane potential, and DNA is introduced through small pores formed on the membrane | Stable transfection Instantaneous transfection All cells | Widely applicable, in addition to plasmids, it can also be transfected into large genomes (>65kb), but the cell lethality is high, and the amount of DNA and cells used is large. Therefore, the electroporation experimental conditions need to be optimized according to different cell types, with copy numbers ranging from 1 to 20 |
This product is only for scientific experimental research and should not be used in clinical diagnosis, treatment, or other fields.
Q:The cells we transmit contain antibiotics. Will antibiotics absolutely affect the transfection efficiency?
A:能不加尽量不加,理论上抗生素进不到细胞内,但是在转染试剂的作用下,细胞的通透性增加,抗生素可能进到细胞内,对实验结果完成影响。
Q:The protocol mentions that Opti MEM should be used to dilute plasmids and transfection reagents. Can't it be diluted with 1640? Why do plasmids and transfection reagents need to be diluted first and then mixed? Can't the transfection reagent be directly added to the plasmid solution and mixed, and then diluted with culture medium?
A:EZ Trans cell transfection solution belongs to the cationic type of transfection reagent, which requires certain conditions to form complexes with DNA. At an appropriate concentration, it is beneficial for cations to encapsulate DNA and form well shaped complexes that enter cells. If the two are directly mixed, they can easily form a polymer with diverse aggregation phases, which may expose DNA on the surface and make it difficult for it to enter cells, thereby reducing transfection efficiency. So the mixed sequence of EZ Trans and DNA is also important. Most cell cultures use fetal bovine serum, which has complex components that no one can explain in terms of composition and content. The composition of the cell transfection solution is relatively clear, but it cannot be determined whether it will bind to unknown components of fetal bovine serum. In fact, this phenomenon can cause adverse effects in most transfection reagents, so the influence of serum should be avoided during the formation of EZ Trans DNA complexes.
Q:The protocol mentions that "detecting transfection efficiency within 24-48 hours" refers to the 24-48 hours after removing the culture medium containing the complex and replacing it with a culture medium containing serum, or does it refer to the 24-48 hours after adding the complex and transfection?
A:All times are counted from the contact between EZ Trans DNA complex and cells.
Q:Low transfection efficiency - no efficient DNA transfection reagent complex was generated
A:Do not use products containing serum when preparing DNA transfection reagent complexes. In most cases, efficient DNA transfection reagent complexes can be obtained using serum-free DMEM culture medium. We recommend using a serum containing medium for transfection, and the single host nan complex must be formed in serum-free medium. Attention: It is best to avoid Opti MEN culture medium even if its serum content is low.
Q:Low transfection efficiency - there are inhibitors that inhibit the formation of DNA transfection reagent complexes during the process of generating DNA transfection reagent complexes
A:Serum free culture medium is crucial for the formation of efficient DNA transfection reagent complexes. During the process of generating complexes, avoid using high concentrations of phosphates, chondroitin sulfate, hyaluronic acid, dextran sulfate, or other sulfated proteoglycans.
Q:Low transfection efficiency - The transfection reagent complex contains serum
A:When preparing DNA capture reagent complexes, please use serum-free medium or Opti MEM culture. In most cases, using serum-free DMEM medium can obtain DNA transfection reagent complexes with improved efficiency.
Q:Low transfection efficiency, high cytotoxicity - cell density not optimized
A:During DNA transfection and DNA siRNA co transfection, the cell density is controlled between 60% -70%; During siRNA transfection, the cell density is controlled at around 50%.
Q:How to determine the transfection efficiency of the experiment?
A:It is recommended to set a positive control, such as green fluorescent protein (GFP), for each transfection.
Q:Low transfection efficiency - leaving the transfection complex at room temperature for a long time
A:The EZ Trans cell transfection reagent should be stored at 4 ℃. The optimal placement reagent and temperature for transfection complexes are 10-20 minutes at room temperature. Prolonged placement can significantly reduce transfection efficiency, so it is important to control the placement time within 20 minutes.
Q:Low transfection efficiency - High speed centrifugation transfection complex
A:When forming transfection complexes, the standard operating procedure recommends vortex mixing transfection reagents and DNA or siRNA within a few seconds to produce efficient transfection complexes. If you want to centrifuge the composite adhered to the pipe wall, be sure to use extremely low speed centrifugation, such as 80g centrifugation for 5 seconds.
Q:High cytotoxicity - excessive use of DNA
A:Suggest drawing a dose-response relationship curve to determine the optimal amount of DNA used.
Q:High cytotoxicity - too much transfection reagent used
A:Suggest drawing a dose-response relationship curve to determine the optimal transfection reagent dosage.
Q:High cytotoxicity - poor cell state during transfection
A:The transfection efficiency of EZ Trans cell transfection reagent is not affected by the serum in the culture medium. Therefore, it is recommended to use a complete culture medium containing 10% fetal bovine serum and antibiotics during the transfection process to improve the cell state.
Q:High cytotoxicity - antibiotics added too early during stable transfection
A:Please add selective antibiotics 48 hours after transfection.
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