Polybrene (hexadimethrine bromide)
Description
- Preparation of recombinant retrovirus stock: Culture cells containing a monolayer of transfection retrovirus packaging cells in a 100 mm dish with 5 mL growth medium (5% serum). After 24 hours, remove the culture medium and filter it through a 0.45 μm filter.
- Culturing cells for infection: In a 100 mm dish, add 10 mL complete growth medium with a cell density of 5×105 per dish.
- Virus infection: After 24 hours of cell culture, remove the complete culture medium. Infect cells with 2 mL virus supernatant containing Polybrene (final concentration: 5-10 μg/mL) at 37°C for 3-6 hours.
- Collect virus particles: Add 8 mL complete growth medium. After 2-3 days of culture, collect the culture medium to obtain virus particles.
Polybrene (hexadimethrine bromide), also known as Polybrene, is a cationic polymer commonly used in DNA transfection experiments with mammalian cells to enhance the transfection efficiency of liposomes. It is widely employed in retrovirus-mediated gene transfection and lentivirus-mediated gene transfection. The mechanism of action may involve neutralizing the sialic acid on the cell surface, facilitating adsorption by overcoming electrostatic repulsion with virus particles. Polybrene is also recognized as an anticoagulant (heparin antagonist) and is frequently used in the production of non-specifically aggregated red blood cells. Additionally, in protein sequencing, low doses of Polybrene have been shown to significantly improve the degradation of peptides in automated sequencing analysis. The addition of Polybrene to PVDF membranes can also enhance membrane affinity.
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