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GeticoFect 293 Transfection Kit Instruction Manual1. IntroductionThe GeticoFect 293 Transfection Kit, developed by Geneticsci Biotech (Shanghai) Co., Ltd., is a specialized and eff
GeticoFect 293 Transfection Kit Instruction Manual1. IntroductionThe GeticoFect 293 Transfection Kit, developed by Geneticsci Biotech (Shanghai) Co., Ltd., is a specialized and eff
Catalog Number | Specification | Price (CNY) |
182601 | For 1L | 8,500.00 |
182602 | For 10L | 76,000.00 |
182603 | For 50L | 344,000.00 |
In the field of modern biotechnology and biopharmaceuticals, efficient cell transfection technology is a key link to achieve important goals such as large-scale production of recombinant proteins and gene function research. GeticoFect™ 293 Transfection Kit is an innovative product carefully developed to meet the transfection needs of 293 cells, aiming to provide researchers and biotech enterprises with an efficient, reliable, and convenient transfection solution.
GeticoFect™ 293 Transfection Kit focuses on the transient transfection of HEK (Human Embryonic Kidney) 293 cells in high-density culture. As a widely used cell line in biopharmaceuticals and genetic research, HEK 293 cells have advantages such as rapid growth, easy cultivation, and good transfectability. However, traditional transfection methods often struggle to achieve ideal transfection efficiency and protein expression levels when dealing with 293 cells in high-density culture.
The high-efficiency transfection reagent and transfection enhancer in this kit are carefully designed to maximize the protein expression level of 293F cells cultured in 293 expression medium. Through a unique formula and mechanism of action, the transfection reagent can quickly and effectively deliver exogenous genes into 293 cells, while the transfection enhancer further promotes the transcription and translation processes of genes, thereby significantly increasing protein expression.
The kit is equipped with a matching transfection enhancer specifically used to improve transfection performance and protein expression, making it particularly suitable for the transfection of high-density suspension cell cultures. Under high-density culture conditions, the interaction between cells is more complex, and the supply of nutrients and oxygen also faces greater challenges. GeticoFect™ 293 Transfection Kit can overcome these difficulties, ensuring efficient expression of exogenous genes in high-density suspension cells and providing strong support for large-scale protein production.
Compared with other transfection reagents used for high-density 293 cell culture, GeticoFect™ 293 Transfection Kit has a significant advantage in protein yield, which can reach 2 to 20 times that of other reagents. This means that researchers and enterprises can obtain higher yields at a lower cost when using this kit for protein expression experiments, greatly improving experimental efficiency and economic benefits.
Using the same transient expression protocol commonly used in current low-density 293 suspension culture systems, users can easily transition from low-density systems to high-yield, high-density 293 expression systems. This compatibility eliminates the need for researchers to re-optimize experimental protocols, reducing the complexity and time cost of experiments, while also ensuring the consistency and reliability of experimental results.
GeticoFect™ 293 Transfection Kit provides robust and reproducible transfection results. In multiple experiments, the kit can maintain stable transfection efficiency and protein expression levels, giving users confidence in the experimental results. Whether in basic research laboratories or large-scale production workshops, reliable experimental results are a key factor in promoting scientific research and production processes.
The kit has excellent adaptability to culture volume, capable of expanding the culture volume from 1 mL to 1000 liters while maintaining the same protein yield per unit volume. This means that whether for small-scale experimental research or large-scale industrial production, users can use this kit for efficient transfection operations to meet the needs of experiments and production of different scales.
GeticoFect™ 293 Transfection Kit consists of cationic nano GeticoFect™ 293 Transfection Reagent and GeticoFect™ 293 Transfection Enhancers 1 and 2. The cationic nano-transfection reagent has good biocompatibility and transfection efficiency, and can form stable complexes with exogenous genes, which are efficiently delivered into cells through electrostatic interaction with the cell membrane.
Transfection Enhancers 1 and 2 are added 15 to 16 hours after transfection, each with a unique mechanism of action. The transfection enhancers can enhance the transfection effect, improve cell activity, and reduce cell damage during the transfection process. At the same time, they can also promote the transcription and translation processes of exogenous genes, further increasing the protein expression level. Through this synergistic effect, the transfection enhancers and transfection reagent work together to ensure that 293 cells can efficiently express exogenous proteins.
In conclusion, with its precise design, outstanding performance, and scientific formula, GeticoFect™ 293 Transfection Kit provides an ideal solution for the transfection and protein expression of 293 cells. Whether exploring the mysteries of genes in the field of scientific research or realizing large-scale protein production in the biopharmaceutical industry, this kit will become a powerful assistant for researchers and enterprises, promoting the continuous development of the biotechnology and biopharmaceutical industries.
Cat. No. | Specification | Price(￥) |
182601 | 1L | 8500.00 |
182602 | 10L | 68000.00 |
182603 | 50L | 289000.00 |
Q: What is the design principle of GeticoFect 293 Transfection Kit, and what are its unique advantages?
A: GeticoFect 293 Transfection Kit is specifically designed for efficient transfection of 293 cell lines. Its principle is based on an optimized cationic liposome formula. These cationic liposomes can form stable complexes with nucleic acids (such as plasmid DNA, siRNA, etc.) through electrostatic interaction. The complexes can effectively bind to the surface of 293 cells, enter the cells through endocytosis, and then release the nucleic acids to achieve gene introduction or interference.
The unique advantages are as follows:
• Ultra-high transfection efficiency: In experiments with various 293 cell lines (such as 293T, HEK293, 293F, 293A, etc.), the transfection efficiency can be as high as over 95%, which is significantly better than many general transfection reagents. For example, in 293T cell transfection experiments, the use of GeticoFect 293 Transfection Kit can make the positive expression rate of target genes much higher than that of similar products, which can greatly meet experiments with strict requirements on transfection efficiency, such as gene function verification and gene introduction in the early stage of recombinant protein production.
• Low cytotoxicity: Cationic liposomes are made of special degradable biological materials, minimizing cytotoxicity. Twenty-four hours after cell transfection, the cell death rate is only about 5%, which allows cells to maintain a good physiological state after transfection, reducing the interference of cytotoxicity on experimental results, and making the experimental data more reliable. It is especially suitable for long-term experiments with high requirements on cell state, such as the construction of stable cell lines.
• Easy operation: The operation process has been simplified and optimized. First, fully mix the transfection reagent with nucleic acid to form a transfection complex, and then directly add the complex to the medium containing cultured 293 cells, without complex pretreatment steps. Moreover, serum does not affect its transfection effect, and transfection can be carried out directly in serum-containing medium. There is no need to deliberately add or replace the culture medium before and after transfection, saving experimental time and labor costs.
Q: Is this kit suitable for all 293 series cells, and can it be used for other cell types?
A: GeticoFect 293 Transfection Kit has wide applicability to 293 series cells, including but not limited to the following common cells:
• 293T cells: As a commonly used cell line for gene function research, the kit can achieve efficient transfection in 293T cells, facilitating experiments such as constructing cell models with gene overexpression or gene knockdown.
• HEK293 cells: Namely human embryonic kidney 293 cells, which themselves have high transfection efficiency. The use of GeticoFect 293 Transfection Kit can further improve the transfection effect, and perform well in the production and research involving 293 cells, such as recombinant protein expression and vaccine development.
• 293F cells: Often used for large-scale suspension culture to produce recombinant proteins. The kit can adapt to its suspension culture characteristics, efficiently complete transfection, and ensure protein yield and quality.
• 293A cells: Widely used in the production of gene therapy vectors and other aspects. The kit can effectively introduce target genes into 293A cells, promoting related research and applications.
However, for other cell types outside the 293 series, although the kit is mainly optimized for 293 cells, some cells may also be applicable. For example, some cells insensitive to cationic liposome transfection may not achieve ideal transfection effects; while certain epithelial cell lines or some primary cells, if their surface characteristics have certain similarities to 293 cells, may achieve low-efficiency transfection after strict pre-experiment optimization of transfection conditions (such as adjusting the ratio of transfection reagent to nucleic acid, transfection time, etc.), but it is not recommended to use it as a routine transfection option. Before attempting to use the kit on non-293 cells, a small-scale pre-experiment must be conducted to evaluate its feasibility.
Q: When using GeticoFect 293 Transfection Kit, what may cause unsatisfactory transfection efficiency, and how to solve it?
A: Unsatisfactory transfection efficiency may be caused by various factors, and the corresponding solutions are as follows:
• Poor cell state: 293 cells should be in the logarithmic growth phase with good viability during transfection. Excessive cell passage times, too high or too low cell density will affect the transfection effect. For adherent-growing 293 cells (such as 293T, HEK293), the cell confluency during transfection is recommended to be controlled at 70%-80%; for suspension-growing 293F cells, the cell density should be adjusted to an appropriate range (generally 1×10^6 - 3×10^6 cells/mL). Thaw cells regularly and avoid using cells with excessive passage times. For example, the passage times of 293T cells should not exceed 30 generations as much as possible.
• Nucleic acid quality issues: The purity, integrity, and concentration of nucleic acids have a significant impact on transfection efficiency. Plasmid DNA should have no obvious degradation, with an OD260/280 ratio between 1.8 and 1.9, and no contamination by impurities such as proteins and RNA; siRNA should avoid prolonged exposure to air or repeated freeze-thawing to prevent degradation. Use high-quality nucleic acid extraction kits, and determine the concentration and purity of nucleic acids before transfection. If the nucleic acid concentration is too low, the amount of nucleic acid can be appropriately increased, but the issue of cytotoxicity must also be considered, and the optimal amount of nucleic acid should be determined through pre-experiments.
• Inappropriate ratio of transfection reagent to nucleic acid: This is one of the key factors affecting transfection efficiency. The optimal ratio of different types and sizes of nucleic acids to transfection reagents varies. For plasmid DNA, it is recommended to try different ratios such as 1:1, 2:1, 3:1 (μg DNA:μL transfection reagent) in pre-experiments to find the ratio with high transfection efficiency and low cytotoxicity; for siRNA, a similar optimization process is required, and generally, adjustments can start from the ratio recommended by the kit.
• Non-standard transfection operation: Details during the operation process can also affect the transfection results. For example, in the preparation of transfection complexes, the transfection reagent and nucleic acid must be fully mixed, but violent oscillation should be avoided; when adding the transfection complex, it should be added dropwise slowly and mixed gently to ensure that the complex is evenly distributed in the cell culture medium. During the transfection process, the cell culture conditions should be kept stable, with the incubator temperature maintained at 37°C and CO₂ concentration at 5%, to avoid fluctuations in temperature and CO₂ concentration affecting cell state and transfection efficiency.
Q: After using the kit, the cytotoxicity is relatively high. What may be the reason, and how to reduce cytotoxicity?
A: High cytotoxicity may be caused by the following reasons, with corresponding solutions:
• Excessive amount of transfection reagent: A high concentration of transfection reagent can be toxic to cells. Cytotoxicity can be reduced by reducing the amount of transfection reagent. On the premise of ensuring a certain transfection efficiency, gradually reduce the amount of transfection reagent (e.g., reduce by 20% each time starting from the recommended amount), and observe the changes in cytotoxicity and transfection efficiency to find the optimal balance between the two.
• Prolonged transfection time: Prolonged incubation of the transfection complex with cells will increase the burden on cells. Perform transfection according to the time recommended in the instruction manual. Generally, 4-6 hours after transfection, fresh serum-containing medium can be replaced to dilute the residual transfection reagent, shorten the contact time between the transfection reagent and cells, and reduce cytotoxicity.
• Cell sensitivity to the reagent: Although the kit has low cytotoxicity to 293 cells, the tolerance of different cell lines or different batches of cells of the same cell line to the transfection reagent may vary. For sensitive cells, they can be cultured in serum-containing medium before transfection to enhance cell resistance; or try adding cell protectants (such as antioxidants like N-acetylcysteine) to the transfection system to reduce cell damage.
• Impure nucleic acid: Impurities in nucleic acids (such as endotoxins, incompletely removed proteins, etc.) may also increase cytotoxicity. Use high-quality nucleic acid extraction methods to ensure that the nucleic acid purity meets the standard. If necessary, further purify the extracted nucleic acid to remove impurities and reduce the risk of cytotoxicity.
Q: How should GeticoFect 293 Transfection Kit be stored, and what is its shelf life?
A: The transfection reagent in the kit should be stored in a 4°C refrigerator; freezing should be avoided, as freezing may damage the liposome structure and affect the transfection effect. Other supporting reagents (such as buffers) should be stored at the temperature required in the instruction manual, generally 4°C. The kit has a shelf life of 12 months when unopened and stored under correct conditions. After opening, it should be used as soon as possible, and attention should be paid to maintaining the sterility of the reagent during use to avoid contamination. When approaching the expiration date, a small-scale pre-experiment can be conducted first to detect whether the performance of the kit has declined. If there are significant changes in indicators such as transfection efficiency and cytotoxicity, a new kit should be replaced in a timely manner.
Q: What precautions should be taken during the operation process?
A: The following precautions should be noted during the operation:
• Aseptic operation: The entire operation should be carried out in a clean bench, using sterile pipette tips, centrifuge tubes, cell culture dishes, and other consumables. Operators should wear sterile work clothes, gloves, and masks to prevent microbial contamination of reagents and cells, and avoid abnormal cell state or transfection failure caused by contamination.
• Reagent preparation: Before use, equilibrate the reagents in the kit to room temperature, and mix gently by inverting, avoiding violent oscillation to generate bubbles, which may affect the activity of the transfection reagent. The transfection reagent and nucleic acid should be avoided from exposure to strong light, especially for samples containing RNA, as light may cause nucleic acid degradation and reduce transfection efficiency.
• Accurate sample addition: Use a high-precision pipette to accurately measure the transfection reagent and nucleic acid to ensure the accurate ratio of transfection reagent to nucleic acid. Add the sample dropwise slowly to avoid generating bubbles. After adding the sample, gently pipette or vortex gently with a pipette to ensure that the transfection complex is fully formed and evenly distributed.
• Record experimental parameters: Record various parameters during the experiment in detail, including cell line name, cell generation, cell density, dosage and ratio of transfection reagent and nucleic acid, transfection time, culture conditions, etc. These records are helpful for the repetition of subsequent experiments and result analysis, and facilitate troubleshooting problems in the experiment.
• Safety protection: Although the kit itself does not involve biosafety risks, when handling transfected cells and waste, operations should be carried out in accordance with laboratory safety regulations. For cells and related waste that may contain recombinant genes, proper treatment (such as autoclaving) should be performed to prevent potential risks caused by gene diffusion.
