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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-3580 is the HeLa-based cell line P4 CCR5+ that stably expresses CD4 and CCR5, as detected by flow cytometry and fluorescence-activated cell sorting (FACS), and have Tat-inducible β-galactosidase activity under the control of human immunodeficiency virus 1 (HIV-1) long terminal repeat (LTR), which is transactivated by HIV or Simian Immunodeficiency Virus (SIV) TAT. Therefore, cells infected with HIV or SIV should express ß-galactosidase.
ARP-3580 was generated by subcloning cDNA encoding CCR5 into the retroviral vector pBABE-puro. Amphotropic virus stocks were prepared by cotransfecting 293T cells with the resulting pBABE-puro construct, a VSV-G envelope expression vector, and pSV-gag-pol.Ψ-env-. Supernatants were collected after 48 hours and used to infect P4 cells. After another 48 hours, cells were selected in medium containing 1 µg per mL puromycin. β-chemokine receptor expression can be maintained by culturing the cells in medium containing 0.5 µg per mL to 1.0 µg per mL puromycin.
ARP-3580 is also referred to as P4.R5 MAGI cells.
Growth Characteristics: Thaw cells quickly at 37°C and immediately place them in a centrifuge tube containing 10 mL culture medium. Centrifuge at 400 × g for 4 minutes. Aspirate the supernatant and resuspend the cells in 10 mL fresh culture medium. Plate the cells into a 10-cm tissue culture dish. The cells are adherent, and can be removed from the culture dish with trypsin. Check the cells the next day to see if they need to be split. The cells require a minimum of 3 to 4 days to recover. A dish split 1:10 should be confluent after three days, and cells should be split at least twice weekly. The cells should grow singly, and should not be allowed to become too clumpy.
Growth Medium: DMEM supplemented with 10% fetal bovine serum and 1.0 µg per mL puromycin
ARP-3580 is negative for bacteria, fungi and Mycoplasma.
Each vial of ARP-3580 contains frozen cells in DMEM supplemented with 20% fetal bovine serum, 1 µg per mL puromycin and 10% dimethyl sulfoxide (DMSO). Please refer to the attached file(s) for more information.
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