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Applications: Generate cell pools or stable cell line expressing membrane-bound TNFα following puromycin selection.Study anti-TNFα antibodies ADCC (antibody dependent cellular cytotoxicity) potential.
Background: Tumor necrosis factor (TNF, also known as TNFα) is a cytokine produced predominantly by activated macrophages and T lymphocytes. It has been identified as a key regulator in inflammatory and immune responses. TNF signaling pathways are triggered by binding to one of two distinct receptors, designated TNFR1 (TNF receptor 1) and TNFR2, which are differentially regulated on various cell types in normal and diseased tissues. TNFα exists in both a trimeric membrane-bound form (mTNFα) and as a soluble protein. TNFα is synthetized in a precursor form, a cell surface type II transmembrane protein, which is cleaved by metalloproteinases such as TACE (TNFα converting enzyme) into a soluble peptide. Soluble TNFα can then bind to its receptors and activate downstream signaling pathways. Transmembrane TNFα can also bind to TNFα receptors and induce cellular responses. For instance, it can enhance cytotoxicity in NK cells, while in the liver it can trigger hepatitis. Anti-TNFα antibodies can bind to mTNFα and trigger antibody-dependent cell-mediated cytotoxicity (ADCC) or complement-dependent cytotoxicity (CDC) to destroy the mTNFα-expressing inflammatory cells, being a promising therapy for inflammatory diseases.
Description: Membrane-Bound TNFα Lentivirus are replication incompetent, HIV-based, VSV-G pseudotyped lentiviral particles that are ready to transduce nearly all types of mammalian cells, including primary and non-dividing cells. These particles result in expression of uncleavable, membrane-bound TNFα (Tumor necrosis factor alpha) driven by an EF1a promoter, and a puromycin selection marker.
Formulation: The lentivirus particles were produced from HEK293T cells. They are supplied in cell culture medium containing 90% DMEM + 10% FBS. Virus particles can be packaged in custom formulations and produced at higher titers by special request, for an additional fee.
Storage Stability: Lentiviruses are shipped with dry ice. For long-term storage, it is recommended to store the lentiviruses 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 freeze/thaw cycles
Biosafety Level: BSL-2
References: Wang F., et al., 2017 Mol Med Rep 16: 1021-1030.
Horiuchi T., et al., 2010 Rheumatology 49(7):1215-1228.
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