2H4 Cell Line
A cell line for IL-2 Luc assay.
2H4 cells are utilized for the in vitro immunotoxicity test (IL-2 Luc Assay), which is approved as an internationally recognized testing protocol by the Organization for Economic Co-operation and Development (OECD). 2H4 cells are derived from human acute T lymphoblastic cell line (Jurkat cells) stably introducing luciferase genes, in which green-emitting luciferase (Stable Luciferase Green, SLG) is expressed under the control of interleukin-2 (IL-2) promoter, orange-emitting luciferase (Stable Luciferase Orange, SLO) is expressed under the control of interferon (IFN)- γ promoter, and red-emitting luciferase (Stable Luciferase Red, SLR) is expressed under the control of GAPDH promoter, used as an internal control reporter. To evaluate immunotoxicity, transcriptional change of IL-2 promoter of human T cells by immunotoxic chemicals is simply measured by luciferase reporter assay.
*2H4 cells were established by Tsuruga Institute of Biotechnology, TOYOBO CO., LTD., and Department of Dermatology, Tohoku University School of Medicine (Saito R. et al.(2011) Toxicol. Appl. Pharmacol., 254, 245-255).
Assay components
Here is the list of assay components.
Reporter cell line: 2H4 Cell Line
2H4 cells were established by Tsuruga Institute of Biotechnology, TOYOBO CO., LTD., and Department of Dermatology, Tohoku University School of Medicine (Saito R. et al.(2011) Toxicol. Appl. Pharmacol., 254, 245-255).
Assay reagent
Tripluc® Luciferase assay reagent (TOYOBO Cat#MRA-301)
Controls
-Ionomycin (CAS:56092-82-1, Sigma Cat#I0634) -Phorbol 12-myristate 13-acetate (PMA) (CAS:16561-29-8, Sigma Cat#P8139) -Ethanol (e.g., Wako Cat#057-00456) -Dimethyl sulfoxide (DMSO) (CAS:67-68-5, Sigma Cat#D5879) -Distilled water (GIBCO Cat#10977-015)
Medium
-RPMI-1640 (GIBCO Cat#11875-093, 500 mL) -FBS (Biological Industries Cat#04-001-1E Lot: 1524129) -Antibiotic-Antimycotic (GIBCO Cat#15240-062) -HygromycinB (CAS:31282-04-9, Invitrogen Cat#10687-010) -G418 (CAS:108321-42-2, WAKO Cat#074-06801) -Puromycin (CAS:58-58-2, InvivoGen Cat#ant-pr-1)
IL-2 Luc assay: OECD test guideline No. 444A
2H4 cells are utilized for the in vitro immunotoxicity test (IL-2 Luc Assay), which is approved as an internationally recognized testing protocol by the Organization for Economic Co-operation and Development (OECD).
To evaluate immunotoxicity, transcriptional change of IL-2 promoter of human T cells by immunotoxic chemicals is simply measured by luciferase reporter assay.
How to obtain cell line
Cell lines must be transfered under MTA (Material Transfer Agreement). Users must enter MTA with Tottori Bioscience Foundation through Biospecimen Laboratories, Inc. (agent) before order and obtain research license.
Order(s) for cell line should be placed after execution of MTA. Pre-payment is necessary.
Costs for initial transfer includes research license for two years.
Biospecimen Laboratories, Inc. will send cell lines and other products ordered by users. Users’ accounts will be used for shipment.
Users can use cell lines following standard protocols.
