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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-1300 is a HeLa derivative that contains a stably integrated copies of fB, a rev-deficient human immunodeficiency virus 1 (HIV-1) HXB2 molecular clone.
The HLfB cell line was generated by cotransfection of HeLa cells with the plasmids fB and pSV2neo. Clone HLfB produces high levels of Gag and Env proteins only in the presence of Rev, and was selected in G418. The rev gene can be introduced into HLfB by transfection with a rev plasmid, by fusion to a rev-expressing cell line, or by fusion to Escherichia coli protoplasts containing bacterially-expressed Rev. High levels of virus are produced after introduction of Rev. Morphology is epithelial-like.
Growth Characteristics: Split cells twice weekly at a ratio of 1:10. HLfB cells are stable and do not need to be maintained in selection medium. If growth in selection medium is desired, propagation medium containing 500 µg per mL G418 should be used. The culture flask should be changed every two weeks. Avoid repeated passages of this cell line in culture because a gradual decrease in Env production occurs after the 15th passage. The available lot is passage #7.
Growth Medium: RPMI 1640 supplemented with 10% fetal bovine serum
ARP-1300 is negative for bacteria, fungi and Mycoplasma.
Each vial of ARP-1300 contains approximately 4 × 106 cells in RPMI 1640 supplemented with 20% fetal bovine serum and 10% dimethyl sulfoxide (DMSO). Please refer to the appropriate data sheet for lot-specific information.
Drysdale, C. M. and G. N. Pavlakis. “Rapid Activation and Subsequent Downregulation of the HIV-1 Promoter in the Presence of Tat: Possible Mechanisms Contributing to Latency.” J. Virol. 65 (1991): 3044-3051.PubMed: 2033665. Felber, B. K., C. M. Drysdale and G. N. Pavlakis. “Feedback Regulation of Human Immunodeficiency Virus Type 1 Expression by the Rev Protein.” J. Virol. 64 (1990): 3734-3741. PubMed: 2196381.
Felber, B. K. and G. N. Pavlakis. “Cell Fusion and Transfection of HL3Ta and HeLa-tat Cell Lines.” Courier 91 (1991): 8-10.
Hadzopoulou-Cladaras, M., et al. “The Rev (Trs/Art) Protein of Human Immunodeficiency Virus Type 1 Affects Viral mRNA and Protein Expression via a cis-Acting Sequence in the env Region.” J. Virol. 63 (1989): 1265-1274. PubMed: 2783738.
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