Retroviral Expression System

Product Discontinued

Applications/Dilutions

• Dilutions:
  • Retroviral Production
• Application Notes: Retrovirus particles are fragile. They are easily inactivated by 0.1% detergent, chloroform, phenol, 1% bleach, 70% ethanol, at 65oC for 30 min, pH <6.5 or >9.0, UV light, and autoclaving. Simple high-speed centrifugation (100,000g x 90 min) produces enough hydrodynamic shear to strip many virions of their gp70, and thus infectivity (although reverse transcriptase activity is preserved). When filter sterilizing retrovirus, be sure to use non-detergent treated 0.22 or 0.45 um filters. Any trace of detergent will strip virus envelope and reduce your titers. Filter before freezing, and not after in order to avoid looses due to aggregation. All murine retrovirus vectors produced in either mouse cells (like the NIH 3T3- based packaging cell lines) or primate cells (COS and 293) are rapidly inactivated by human serum complement, with kill kinetics of 2-3 logs in 5 min at 37C. Human C1q initiates the cascade by binding p15ETM at the virion surface. This is an antibody-independent process. Murine retroviruses are heat labile. They have an infectious half-life of only 6 h in culture medium at 4C. Use in retroviral production reported in scientific literature (PMID: 24312621)

Reactivity Notes

Human and mouse reactivity reported in scientific literature (PMID: 24312621)

Packaging, Storage & Formulations

• Storage: Storage is content dependent.

Notes

Additional Materials Required:
293 Cells may be ordered from ATCC
G418
Cell culture reagents: DMEM, Fetal bovine serum, penicillin-streptomycin
Trypsin-EDTA
Phosphate buffered saline
Cloning cylinders (for picking up stably transfected colonies)
Polybrene for retroviral infection.
Please note that the sequence information for these plasmids is not available.

Alternate Names for Retroviral Expression System

• RetroMax
• Retroviral System

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Support products are guaranteed for 6 months from date of receipt.

Publications for Retroviral Expression System (NBP2-29499)(11)

Publications using NBP2-29499

• Tamaru T, Hattori M, Honda K et al. CRY Drives Cyclic CK2-Mediated BMAL1 Phosphorylation to Control the Mammalian Circadian Clock. PLoS Biol. 2015-11-01 [PMID: 26562092] (RetVir, Mouse)
• Tamaru T, Hattori M, Ninomiya Y et al. ROS Stress Resets Circadian Clocks to Coordinate Pro-Survival Signals. PLoS One 2013-12-02 [PMID: 24312621] (Ret Vir, Human, Mouse)
• Narala SR, Allsopp RC, Wells TB et al. SIRT1 acts as a nutrient-sensitive growth suppressor and its loss is associated with increased AMPK and telomerase activity. Mol Biol Cell. 2008-03-01 [PMID: 18184747] (Human)
  
  Details:
  Packaging plasmids were transfected into 293T cells.
• Silverman MA, Shoag J, Wu J et al. Disruption of SLP-76 interaction with Gads inhibits dynamic clustering of SLP-76 and FcepsilonRI signaling in mast cells. Mol Cell Biol. 2006-03-01 [PMID: 16479002] (Mouse)
• Flamant S, Kortulewski T, Dugray A et al. Osteopontin is upregulated by BCR-ABL. Biochem Biophys Res Commun. 2005-08-12 [PMID: 15993098]
• Hirayama J, Kaneko M, Cardone L et al. Analysis of circadian rhythms in zebrafish. Methods Enzymol. 2005-01-01 [PMID: 15817288] (Zebrafish)
• Xiong Y, Miyamoto N, Shibata K et al. Short-chain fatty acids stimulate leptin production in adipocytes through the G protein-coupled receptor GPR41. Proc Natl Acad Sci U S A. 2004-01-27 [PMID: 14722361] (Mouse)
• Cvijic ME, Xiao G, Sun SC. Study of T-cell signaling by somatic cell mutagenesis and complementation cloning. J Immunol Methods. 2003-07-01 [PMID: 12957416] (Human)
  
  Details:
  pCL-Ampho Packaging Vector (10046P) which is also a component of the RetroMax System (10040K): Fig 2 (293 cells were used as the packaging cell line). Note: A VSV-G Vector, encoding the G glycoprotein of the vesicular stomatitis virus under the control of the CMV immediate-early promoter was used in conjunction with pCL-Ampho to help increase infection efficiency (see pages 297-298 and Fig 2 for details).
• Gao P, Zhou XY, Yashiro-Ohtani Y et al. The unique target specificity of a nonpeptide chemokine receptor antagonist: selective blockade of two Th1 chemokine receptors CCR5 and CXCR3. J Leukoc Biol. 2003-02-01 [PMID: 12554804] (Mouse)
• He KL, Ting AT. A20 inhibits tumor necrosis factor (TNF) alpha-induced apoptosis by disrupting recruitment of TRADD and RIP to the TNF receptor 1 complex in Jurkat T cells. Mol Cell Biol. 2002-09-01 [PMID: 12167698] (Human)
• Ronnov-Jessen L, Villadsen R, Edwards JC et al. Differential expression of a chloride intracellular channel gene, CLIC4, in transforming growth factor-beta1-mediated conversion of fibroblasts to myofibroblasts. Am J Pathol. 2002-08-01 [PMID: 12163372] (Mouse)

FAQs for Retroviral Expression System (NBP2-29499). (Showing 1 - 1 of 1 FAQs).

1. For Retroviral Expression Systems (#NBP2-29499), could you let us know the composition of Retromax Transfection Buffer? If it is proprietary, we would just like to confirm that no toxic preservatives (e.g. thimerosal and sodium azide) are contained.
   • The Retromax Transfection Buffer contains HEPES, Na2HP04 and NaCl in an optimized proprietary formulation. It does not contain any preservatives. I confirm that no thimerosol or sodium azide is present.