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Application: Screen for activators or inhibitors of NFAT signaling pathways in transduced target cells. Generate NFAT Luciferase-RFP Reporter stable cell line (hygromycin or puromycin resistant).
Background: NFAT (Nuclear factor of activated T-cells) is a family of transcription factors that has an important function in immune responses, for example by inducing the expression of various cytokines (such as IL-2-3-4 and TNF-alpha) in T cells. Members of the NFAT family have been found in many tissue types including heart, skeletal muscle and brain cells. Through their role in the immune system, NFATs are involved in inflammation and these transcription factors are considered promising therapeutic targets for a variety of diseases. NFAT is regulated by Ca2+ and the Ca2+/calmodulin-dependent serine phosphatase, calcineurin. NFAT proteins are phosphorylated and reside in the cytoplasm in resting cells; upon stimulation, they are dephosphorylated by calcineurin, translocate to the nucleus, and induce gene expression.
Description: The NFAT Luciferase-RFP Reporter Lentiviruses are replication incompetent, HIV-based, VSV-G pseudotyped lentiviral particles that are ready to transduce almost all types of mammalian cells, including primary and non-dividing cells. The particles contain a firefly luciferase and RFP (Red Fluorescent Protein) cassette driven by the NFAT response element located upstream of the minimal TATA promoter and a hygromycin or puromycin selection gene to generate stable clones. After transduction, activation of the NFAT signaling pathway in the target cells can be monitored by measuring the luciferase activity or RFP expression. RFP fluoresces red-orange when excited; it has an excitation wavelength of 553 nm, and an emission wavelength of 574 nm.
Formulation: The lentivirus particles were produced from HEK293T cells. They are supplied in cell culture medium containing 90% DMEM + 10% FBS.
Storage Stability: Lentiviruses are shipped with dry ice. For long term storage, it is recommended to store the virus at -80°C. Avoid repeated freeze-thaw cycles. Titers can drop significantly with each freeze-thaw cycle.
Supplied As: Two vials (500 µl x 2) of lentivirus at a titer >107 TU/ml. The titer will vary with each lot; the exact value is provided with each shipment.
Warnings: Avoid repeated freeze/thaw cycles.
Biosafety Level: BSL-2
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