Frequently requested material


C7-50HCV coreMouse IgG1Affinity BioReagents MA1-080Virology 1996;222:51
1B6HCV NS3Mouse IgG1Axxora ALX-803-059-R100J Virol 2000;74:2293
5B-3B1HCV NS5BMouse IgG2bAxxora ALX-803-060-R100J Biol Chem 2002;277:593
5B-12B7HCV NS5BMouse IgG2aAxxora ALX-803-061-R100J Biol Chem 2002;277:593

Cell lines

Our laboratory has established a comprehensive panel of U-2 OS human osteosarcoma-derived cell lines inducibly expressing HCV proteins. Requests should be addressed in written, with a brief description of the planned research project and confirmation that these cell lines will be used only for non-commercial research purposes in your laboratory. Where applicable (see list below), please contact also Dr. Charles M. Rice at The Rockefeller University (Contact) for an MTA for the HCV H prototype (J Virol 1993;67:1385) or consensus cDNA clones (Science 1997;277:570).

UTHCHCV core (gt 1b)Unpublished
UTHHCV 5'NCR-core-part E1 (gt 1b, aa 1-326)Virology 1996;222:51
UTHLHCV 5'NCR-part core-luc (gt 1b, aa 1-82)Virology 1996;222:51
UTHCNS2HCV 5'NCR-core-part NS2 (gt 1b, aa 1-1134)Unpublished
UTHCNS3HCV 5'NCR-core-part NS3 (gt 1b, aa 1-1523)BBRC 1998;246:920
UHCVHCV polyprotein (HCV H prototype clone)Hepatology 1998;28:192
UHCVconHCV polyprotein (HCV H consensus clone)J Biol Chem 2001;276:44052
UCconHCV core (HCV H con)Unpublished
UE1E2conHCV E1E2 (HCV H con)Unpublished
UCp7conHCV core-E1E2-p7 (HCV H con)Unpublished
UNS3-5BconHCV NS3-NS5B (HCV H con)Unpublished
UNS2conHCV NS2 (HCV H con)Unpublished
UNS2-3conHCV NS2-part NS3 (HCV H con, aa 810-1227)Unpublished
UNS3P201HCV part NS3 (HCV H prototype, aa 1027-1227)J Virol 2000;74:2293
UNS3-4AHCV NS3-4A (HCV H prototype)J Virol 2000;74:2293
UNS4AconHCV NS4A (HCV H con)Unpublished
UNS4BconHCV NS4B (HCV H con)Virology 2001;284:70
UNS5AconHCV NS5A (HCV H con)J Biol Chem 2002;277:8130
UNS5BconHCV NS5B (HCV H con)J Biol Chem 2001;276:44052
UGFPGFPJ Biol Chem 2001;276:44052

We maintain these cell lines in Dulbecco's Modified Eagle Medium supplemented with 10% heat-inactivated FBS, 2 mM glutamine, 500 µg/ml G418, 1 µg/ml puromycin and 1 µg/ml tetracycline. Media containing tetracycline should be kept protected from light. We split the cells 1:15 on a weekly basis. For maintenance purposes, cells are always cultured in medium containing tetracycline. To derepress the cells, i.e., to induce HCV protein expression, we rinse them twice with PBS. This should be done with careful aspiration of all media and washes since as little as 0.01 µg/ml tetracycline will still markedly repress the activity of the tTA-dependent promoter. We suggest that you always split the cells into medium containing tetracycline and let them settle and become about 80-90% confluent before you derepress them. After tetracycline withdrawal, HCV proteins become detectable by immunoblot after 4-6 h and expression reaches a steady state after 24(-48) h. We freeze the cells in 50% of the above-mentioned medium, 40% FBS, and 10% DMSO at a density of 1-5 Mio cells/ml.

 Last updated on 06/06/2018 at 15:39