This product is specifically developed for the large-scale production of lentiviral vectors (LV). It is an optimized, linearized polyethylenimine (Linear PEI).

Synthesized entirely through chemical processes, this product is free of animal-derived components and exhibits low cytotoxicity. It is provided as a sterile liquid solution and is fully compatible with both suspension and adherent HEK293 cell systems for LV packaging. Featuring high DNA loading capacity, a low transfection complex ratio, and high complex stability, it greatly facilitates large-scale LV production.

Specifications

Components

Storage

This product should be stored at 2~8℃ for 2 years.

Notes

1. This product is for research use only.

2. Please operate with lab coats and disposable gloves, for your safety.

Instructions

HEK293 Suspension Cell Transfection Protocol

1. Cell Preparation

Select an appropriate seeding density based on cell status. It is recommended to adjust the cell density to 2 × 10⁶ cells/mL 24 hours prior to transfection, and to 3.5 × 10⁶ cells/mL at the time of transfection. To enhance viral yield, 5 mM sodium butyrate can be added to the cell suspension either at the time of transfection (before adding the transfection complexes) or 24 hours post-transfection.

2. Transfection Complex Preparation (Example for a 30 mL system)

2.1 Plasmid-to-Reagent Ratio: The ratio of plasmid (μg) to transfection reagent (μL) is 1:3.

2.2 Plasmid Dilution: Dilute 30 μg of plasmid in 3 mL of serum-free medium and mix gently.

2.3 Reagent Addition: Add 90 μL of UltraLV Transfection Reagent to the diluted plasmid solution and mix gently.

2.4 Complex Incubation: Incubate at room temperature for 15 minutes to allow the formation of plasmid-PEI complexes before use.

3. Cell Transfection

3.1 Directly add the plasmid-PEI complexes evenly into the cell culture.

3.2 Continue culturing the cells at 37°C with 5% CO₂, and harvest the virus 48–72 hours post-transfection.

Table 1. Cell Transfection Parameters (For Reference Only)

HEK293 Adherent Cell Transfection Protocol

1. Cell Seeding

Select an appropriate seeding density based on cell status. It is recommended to seed cells at a density of 4 × 10⁶ cells per 10 cm dish, aiming for approximately 70% confluency at the time of transfection the following day.

2. Transfection Complex Preparation (Example for a 10 cm cell culture dish)

1)Plasmid-to-Reagent Ratio: The ratio of plasmid (μg) to transfection reagent (μL) is 1:3.

2)Plasmid Dilution: Dilute 20 μg of plasmid in 500 μL of serum-free medium and mix gently.

3)Reagent Addition: Add 60 μL of UltraLV Transfection Reagent to the diluted plasmid solution and mix by gentle vortexing.

4)Complex Incubation: Incubate at room temperature for 15 minutes to allow the formation of plasmid-PEI complexes before use.

3. Cell Transfection

1)Directly add the plasmid-PEI complexes evenly into the cell culture.

2)Continue culturing the cells at 37°C with 5% CO₂, and harvest the virus 48–72 hours post-transfection.

Documents:

Safety Data Sheet

40825_MSDS_HB260721

Manual

40825_Manual_HB20260721