InfectinTM enhances lentiviral transduction of Hep G2 cells:
Hep G2 cells were transduced with lenti-GFP vector in the presence or absence of Infectin
$150.00 – $2,500.00
Infectin serves as a viral infection enhancer and increases infectivity rates by 5 to 20 fold.
Infectin is a viral infection enhancer designed to facilitate viral penetration of the cortical actin barrier, thereby greatly enhancing productive viral infection. Infectin can be used to facilitate the infection of a variety of host cells by different viruses and viral vectors. Infectin can enhance viral infection rates by 5 to 20 fold. Virongy developed Infectin based on the scientific theory that the actin cytoskeleton is a natural barrier for viral entry and post-entry intracellular migration (Yoder et al., Cell, 2008, 134:782).
Applications
Important
(*The degree of enhancement is affected by the type of virus and cells. Enhancement is strongest for enveloped viruses entering cells via membrane fusion.)
Example 1 – Infectin used to enhance lentiviral infection of suspension CEM-SS cells:
(See Table 2 for scale-up recommendations)
1. Count cells to be infected, and pellet cells by centrifugation at 300 x g for 5 minutes.
Note: Cell viability should be ≥ 80%.
2. Resuspend cells in complete media at a concentration of ~2 x 106 cells mL-1.
3. Use 100 μl of cells (~2 x 105) per infection.
4. Pre-treat cells by adding 10 μl of Infectin so that Infectin concentration is ~1X. Mix and incubate for 30–60 minutes.
For best results, when using Infectin for the first time, start with a higher concentration (final Infectin concentration may be 2X or 3X). The lowest effective dosage can be determined through a titration experiment (final Infectin concentration may be as low as 1X).
5. Add virus to the cells and mix. Note volume of virus used.
6. Add Infectin in an amount equal to 1/10 of the virus volume used, e.g., if 100 μl of virus is used, add 10 μl of Infectin. Incubate at 37°C for 2-4 hours.
7. Add 1 ml fresh complete media to wash cells. Pellet cells as above and remove supernatant. Optional: Repeat once for a total of 2 washes.
8. After washing, resuspend cells in 1 ml complete medium.
9. Culture infected cells as usual to quantify viral replication.
Example 2 – Infectin enhances lentiviral transduction of adherent HDFn cells:
1. Count HDFn (Human Dermal Fibroblast, neonatal) cells to be infected and seed ~5 x 105 cells per well into 6-well plates (2 ml per well).
2. Culture cells until cells stably adhere to the plates (4–12 hours).
Note: Cell viability should be ≥ 80%.
3. Before infection, wash cells with 2 ml medium, and leave 0.5 ml medium in each well.
4. Pre-treat cells by adding 50 μl of Infectin so that Infectin concentration is ~1X. Mix and incubate for 30–60 minutes.
For best results, when using Infectin for the first time, start with a higher concentration (final Infectin concentration may be 2X or 3X). The lowest effective dosage can be determined through a titration experiment (final Infectin concentration may be as low as 1X).
5. Add virus to cells and mix. Note volume of virus used.
6. Add Infectin in an amount equal to 1/10 of the virus volume used, e.g., if 100 μl of virus is used, add 10 μl of Infectin. Incubate at 37°C for 2–4 hours.
7. Add 2 ml fresh media to wash cells.
8. After washing, add 2 ml fresh complete medium.
9. Culture infected cells as usual.
Selected publications from our users