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-100°C or colder, preferably in the vapor phase of a liquid nitrogen freezer
Quantity limit per order for this item is 1. This item can be ordered twice a year. Orders over this limit will be sent to NIAID for approval before shipment. ARP-1294 is a HeLa derivative that contains stably integrated copies of the human immunodeficiency virus 1 (HIV-1) molecular clone HXB2/3gpt. It was generated by cotransfection of HeLa cells with the plasmids pHXB2/3gpt, pSV2neo and selected in G418. Clone HL2/3 was selected based on the high-level production of Gag, Env, Tat, Rev, and Nef proteins. HL2/3 cells do not produce detectable viral reverse transcriptase (RT) or detectable amounts of mature virions. Cocultivation of ARP-1294 with CD4-producing cell line HLCD4-CAT (ARP-1299) results in efficient cell fusion within 6-12 hours. Upon fusion, Tat produced by the HL2/3 cells activates chloramphenicol acetyltransferase (CAT) gene expression in HLCD4-CAT cells. The fusion efficiency can be quantitated by assaying for CAT gene activation. Fusion inhibitors decrease CAT enzyme levels in a dose-dependent manner. ARP-1294 can also be used for the production of Env and other viral proteins in the absence of an infectious virus. ARP-1294 cells should be split 1:10 twice weekly. The cells are stable and do not need to be maintained in the selection medium. If growth in the selection medium is desired, a propagation medium containing 500 μg/mL geneticin (G418) should be used. The culture flask should be changed every two weeks. Repeated passages of this cell line in culture should be avoided because a gradual decrease in Env production occurs after the 15th passage. The recommended propagation medium is 90% DMEM with 10% fetal bovine serum. Each vial of ARP-1294 contains approximately 3.8 × 106 cells (passage # 12) in 0.8 mL of freeze medium. Post-thaw viability was 92%.
Ciminale, V., et al. “A Bioassay for HIV-1 Based on Env-CD4 Interaction.” AIDS Res. Hum. Retroviruses 6 (1990): 1281-1287. PubMed: 2078409.
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