Retrovirus Transduction Kit

Catalog No. ABIN2344940

Product Details

Application: Transfected Cell Culture (TCC)

Brand: ViraDuctin™

Sample Type: Cell Extracts, Cell Samples

Characteristics: The ViraDuctin™-retrovirus complexes will also quickly sediment onto target cells resulting in fast delivery of the viruses than simple diffusion. Both removal of transduction inhibitors and quick delivery of viruses to cells result in higher gene transfer. ViraDuctin™ Retrovirus Transduction Kit provides the following advantages: • Higher transduction efficiency in many cell types compared to reagents such as Polybrene®. • Rapid purification and removal of transduction inhibitors. • Quick delivery of viruses to cells. • Ideal for transduction of nonpermissive cells such as primary cells and stem cells.

Components:

1. ViraDuctin™ Retrovirus Transduction Reagent A (100X) : One sterile tube - 200 μL
2. ViraDuctin™ Retrovirus Transduction Reagent B (100X) : One sterile tube - 200 μL
3. ViraDuctin™ Retrovirus Transduction Reagent C (8X) : Two sterile tubes - 1.5 mL each

Material not included:

1. Retroviral Stock Solution
2. Transfection Reagent
3. Cells and cell culture growth medium
4. 37 °C Incubator

Application Details

Application Notes: Optimal working dilution should be determined by the investigator.

Comment:

• Proprietary reagent cocktail forms supercomplex with retrovirus
• Increases retrovirus transduction efficiency 2- to 6-fold compared to Polybrene®
• Useful for infection of non-permissive cells, including many primary cells and stem cells

Restrictions: For Research Use only

Handling

Precaution of Use: Remember that you will be working with samples containing infectious virus. Follow the recommended NIH guidelines for all materials containing BSL-2 organims.

Storage: 4 °C

Storage Comment: Store all kit components at 4°C until their expiration dates. DO NOT FREEZE.

References

Gandhi, Evdokimova, T Cuenco, Nikiforova, Kelly, Stringer, Bakkenist, Nikiforov: "Homologous chromosomes make contact at the sites of double-strand breaks in genes in somatic G0/G1-phase human cells." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 109, Issue 24, pp. 9454-9, (2012) (PubMed).

Miyoshi, Ishii, Nagai, Hoshino, Mimori, Tanaka, Nagano, Sekimoto, Doki, Mori: "Defined factors induce reprogramming of gastrointestinal cancer cells." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 107, Issue 1, pp. 40-5, (2010) (PubMed).

Target Details

Background: Retroviral gene transfer is a technique for efficiently introducing stable, heritable genetic material into the genome of any dividing cell type. Replication-incompetent retrovirus is usually produced through transfection of the retroviral vector into a packaging cell line. Retroviruses are classified according to the receptors used to enter host cells. Ecotropic virus can recognize a receptor found on only mouse and rat cells. Amphotropic virus recognizes a receptor found on a broad range of mammalian cell types. Retrovirus usually has lower titer than lentivirus which leads to poor gene transfer. The rate at which retroviral vectors bind to and infect cells is mostly controlled by diffusion. During retrovirus infection, only a small fraction of the retroviral vectors can transduce target cells. Virion adsorption is the limiting step of this process. Another cause of poor gene transfer of retrovirus is the presence of transduction inhibitors in retroviral supernatants such as proteoglycans and glycosaminoglycans. The use of polycations, such as Polybrene®, is standard in many retroviral infection protocols owing to the observations of improved infection efficiency by neutralizing the electrostatic repulsion between virion and cell membranes, but the transduction inhibitors still remain in the retroviral supernatant during infection. ViraDuctin™ Retrovirus Transduction Kit is a proprietary formulation for the rapid purification of retrovirus from inhibitors of transductions.