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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-1115 is the HeLa derivative HL3T1, a sensitive indicator cell line for the human immunodeficiency virus 1 (HIV-1) Tat transactivator protein. HL3T1 cells contain stably integrated, silent copies of the HIV-1 long terminal repeat (LTR) promoter linked to the chloramphenicol acetyl transferase (CAT) gene. They produce CAT enzyme upon introduction of active Tat. This cell line was generated by cotransfection of pL3CATt and the plasmid pSV2neo and selected in G418. Clone HL3T1 was selected based on high activation by Tat protein. In addition, CAT expression by HL3T1 is activated by Herpes virus infection. Cell lines activated only by Tat and not Herpes are available from the contributor on request. Morphology is epithelial-like. Contains LTR sequences to +80 in the R region. Contains the entire U3 region, but lacks U5 sequences.
Growth Characteristics: Split cells twice weekly at a ratio of 1:10 just upon reaching confluency, approximately 2.5 × 107 cells per T-75 flask. The culture flask should be changed every two weeks. The cells are considered to be unhealthy when they become spindle-shaped. HL3T1 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.
Growth Medium: DMEM high-glucose supplemented with 10% fetal bovine serum
ARP-1115 is negative for bacteria and Mycoplasma.
Each vial of ARP-1115 contains approximately 4 × 106 cells in DMEM supplemented with 20% fetal bovine serum and 10% dimethyl sulfoxide (DMSO). Please refer to the appropriate data sheet for lot-specific information.
Felber, B. K. and G. Pavlakis. “A Quantitative Bioassay for HIV-1 Based on Trans Activation.Science 239 (1988): 184-187. PubMed: 3422113. Pavlakis, G. N., B. K. Felber and C. M. Wright. “A Fusion Assay for the Detection of HIV Infected Cells.” Human Retroviruses, Cancer and AIDS: Approaches to Prevention and Therapy. (1986) D. Bolognesi (ed.). Alan R. Liss, Inc., New York.
Wright, C. M., et al. “Expression and Characterization of the Trans-Activator of HTLV-III/LAV Virus.” Science 234 (1986): 988-992. PubMed: 3490693.
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