Hieff Trans™ in vitro siRNA/miRNA Transfection Reagent is a PEI cationic, amphiphilic transfection reagent developed for the transfection of siRNA into mammalian cells. Suitable for transfection of siRNA and miRNA. This transfection reagent encapsulates siRNA or miRNA to form a cationic complex, which interacts with negatively charged proteoglycans on the cell surface and adsorbs on the cell surface, enters the cell through endocytosis, and forms an inclusion body. The transfection reagent acts as a proton sponge, which can absorb H+ in the lysosome, and the continuous influx of protons causes the complex to swell and rupture, thereby releasing the exogenous siRNA or miRNA into the cytoplasm.
This product can achieve over 90% expression efficiency of 1 nM siRNA in a wide range of cell lines, avoiding off-target effects. Suitable for transfection of a variety of cells, including Hela, MCF-7, HepG2, CHO, and other adherent cells; and difficult to transfect suspension cell lines, such as K562 or THP-1 cells, can achieve 80% silencing efficiency; Including some primary cells, primary human fibroblasts, and primary human hepatocytes, etc., the silencing efficiency of 80% can be achieved.
- Low toxicity: the transfected cells still maintain good activity.
- Wide adaptability: ordinary cells and primary cells difficult to transfect are fully covered.
- Cell Transfection
|Physical and chemical Properties||Information|
|Cell Type||Established Cell Lines|
|Transfection Technique||PEI-Based Transfection|
|40806||Hieff Trans™ in vitro siRNA/miRNA Transfection Reagent||0.1 mL||0.5 mL||1 mL|
The product should be stored at 2-8ºC for one year. Do not freeze!
- Demonstration of Transfection Effect
Figure 1. The transfection efficiency of Hieff Trans™ in vitro siRNA/miRNA Transfection Reagent was shown above.
(1) Q: What is the difference between DNA transfection and siRNA transfection?
A: 1. Since there are only 20 base pairs, siRNA is much smaller than DNA with thousands or even more than 10,000 base pairs.
2. The transfection reagents and methods used to transfect DNA are not suitable for transfection of siRNA.
3. siRNA transfection has stricter requirements on the cytotoxicity of transfection reagents than DNA transfection.
(2) Q: Does the transfection reagent need to be changed after transfection?
A: For the medium change, two situations can be distinguished:
1. If there is no medium change before transfection, the medium should be changed about 6 hours after transfection to ensure the nutrients needed for cell growth;
2. If there is a change before transfection The medium can be changed as usual when the medium is nutrient deficient.
(3) Q: What is the efficiency of PEI transfection reagent for transfecting siRNA and plasmid?
A: This PEI transfection reagent is specially developed for siRNA, and plasmid transfection is not recommended.
(4) Q: Can the transfection reagent be frozen?
A: Do not freeze, because the transfection reagent is a PEI cationic transfection reagent. Freezing at low temperature will destroy the activity of the PEI transfection reagent. Therefore, it is best to store it at 4 degrees to maintain the best transfection efficiency.
(5) Q: How to improve the transfection efficiency?
A: a: Density of cells at the time of transfection, 30%-50%.
b: Opti-MEM serum-free medium should be used for siRNA and liposome dilution during transfection.
c: The medium can be changed 4-6h after transfection.
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