DESCRIPTION

• Notes: To generate the NFAT luciferase-eGFP reporter stable cell line, remove the growth medium 48 hours after transduction and replace it with fresh growth medium containing the appropriate amount of puromycin for antibiotic selection of transduced cells. The following Lentivirus Reporter Controls are available from BPS Bioscience to meet your experimental needs: Negative Control Luciferase Lentivirus (BPS Bioscience #79578): Ready-to-transduce lentiviral particles expressing firefly luciferase under the control of a minimal promoter. The negative control is important to establish the specificity of any treatments and to determine the background reporter activity. Renilla Luciferase Lentivirus (G418 or Puromycin) (BPS Bioscience #79565): Ready-to-transduce lentiviral particles expressing Renilla luciferase under the CMV promoter. The Renilla Luciferase Lentivirus can serve as an internal control to overcome sample-to-sample variability when performing dual-luciferase reporter assays. Firefly Luciferase Lentivirus (G418, Hygromycin and Puromycin) (BPS Bioscience #79692-G, #79692-H, #79692-P): Ready-to-transduce lentiviral particles expressing firefly luciferase under the CMV promoter. It serves as a positive control for transduction optimization studies. Biosafety: The lentiviruses are produced with the third generation SIN (self-inactivation) lentivector which ensures self-inactivation of the lentiviral construct after transduction and after integration into the genomic DNA of the target cells. None of the HIV genes (gag, pol, rev) will be expressed in the transduced cells, as they are expressed from packaging plasmids lacking the packing signal and are not present in the lentivirus particle. Although the pseudotyped lentiviruses are replication-incompetent, they require the use of a Biosafety Level 2 facility. BPS Bioscience recommends following all local federal, state, and institutional regulations and using all appropriate safety precautions. Troubleshooting Guide: Visit bpsbioscience.com/lentivirus-faq for detailed troubleshooting instructions. For all further questions, please email support@bpsbioscience.com.
• Shiptemp: -80°C (dry ice)
• Warnings: Avoid freeze/thaw cycles.
• Category: Cell Signaling/Lentivirus
• 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-eGFP 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 eGFP cassette driven by the NFAT response element located upstream of the minimal TATA promoter (Figure 1) and a 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 eGFP expression.
• Formulation: The lentivirus particles were produced from HEK293T cells. They are supplied in cell culture medium containing 90% DMEM + 10% FBS.
• 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.
• Unspsc Code: 41106621
• Unspsc Name: Virus mediated expression vectors or kits
• Applications: Screen for activators or inhibitors of NFAT signaling pathways in transduced target cells. Generate NFAT Luciferase-eGFP Reporter stable cell line (puromycin resistant).
• Product Type: Lentivirus
• Biosafety Level: BSL-2
• Related Products: 71274-1, 60690, 60184, 78320
• Storage Stability: Lentiviruses are shipped with dry ice. For long-term storage, it is recommended to store the lentiviruses at -80°C. 
• Scientific Category: Cell Signaling Pathway