Testing Data
(Fig-1: Induction of TNF-alpha promoter activity by phorbol 12-myristate 13-acetate in TNF-alpha Leeporter HEK293 cells.)
TNF-alpha cell line
TNF-alpha Leeporter HEK293 Cell Line
*Monitor the TNF-alpha induction activity.
*Screen for activators or inhibitors of the TNF-alpha signaling pathway.
Culture conditions:
Cells should be grown at 37 degree C with 5% CO2 using DMEM medium (w/L-Glutamine, 4.5g/L Glucose and Sodium Pyruvate) supplemented with 10% heat-inactivated FBS and 1% Pen/Strep, plus 3 ?g/ml of Puromycin.
It is recommended to quickly thaw the frozen cells upon receipt or from liquid nitrogen in a 37 degree C water-bath, transfer to a tube containing 10 ml of growth medium without Puromycin, spin down cells, resuspend cells in pre-warmed growth medium without Puromycin, transfer resuspended cells to T25 flask and culture in 37 degree C-CO2 incubator.
Leave the T25 flask in the incubator for 2~4 days without disturbing or changing the medium until cells completely recover viability and become adherent. Once cells are over 90% adherent, remove growth medium and passage the cells through trypsinization and centrifugation. At first passage, switch to growth medium containing Puromycin. Cells should be split before they reach complete confluence.
To passage the cells, detach cells from culture vessel with Trypsin/EDTA, add complete growth medium and transfer to a tube, spin down cells, resuspend cells and seed appropriate aliquots of cells suspension into new culture vessels. Subcultivation ration = 1:10 to 1:20 weekly. To achieve satisfactory results, cells should not be passaged over 16 times.
1. Harvest TNF-alpha Leeporter™ – HEK293 cells and seed cells into a white solid-bottom 96-well microplate in 100 ul of growth medium at 5 x 10^4 cells/well.
2. Incubate cells at 37 degree C in a CO2 incubator for overnight.
3. The next day, stimulate cells with different concentrations of phorbol 12-myristate 13-acetate (PMA).
4. Incubate at 37 degree C in a CO2 incubator for 6-16 hours.
5. Add 50 ul of luciferase assay reagent (Cat #MBS668917) per well.
6. Incubate at room temperature for 1-5 minutes and measure luminescence using a microplate luminometer.
Testing Data
(Fig-1: Induction of TNF-alpha promoter activity by phorbol 12-myristate 13-acetate in TNF-alpha Leeporter HEK293 cells.)
Testing Data
(Fig-1: Induction of TNF-alpha promoter activity by phorbol 12-myristate 13-acetate in TNF-alpha Leeporter HEK293 cells.)
NCBI and Uniprot Product Information
NCBI Description
This gene encodes a multifunctional proinflammatory cytokine that belongs to the tumor necrosis factor (TNF) superfamily. This cytokine is mainly secreted by macrophages. It can bind to, and thus functions through its receptors TNFRSF1A/TNFR1 and TNFRSF1B/TNFBR. This cytokine is involved in the regulation of a wide spectrum of biological processes including cell proliferation, differentiation, apoptosis, lipid metabolism, and coagulation. This cytokine has been implicated in a variety of diseases, including autoimmune diseases, insulin resistance, and cancer. Knockout studies in mice also suggested the neuroprotective function of this cytokine. [provided by RefSeq, Jul 2008]
Uniprot Description
TNF-a: Cytokine that binds to TNFRSF1A/TNFR1 and TNFRSF1B/TNFBR. It is mainly secreted by macrophages and can induce cell death of certain tumor cell lines. It is potent pyrogen causing fever by direct action or by stimulation of interleukin-1 secretion and is implicated in the induction of cachexia, Under certain conditions it can stimulate cell proliferation and induce cell differentiation. Homotrimer. Interacts with SPPL2B. Belongs to the tumor necrosis factor family.
Protein type: Apoptosis; Cytokine; Membrane protein, integral; Motility/polarity/chemotaxis
Chromosomal Location of Human Ortholog: 6p21.33
Cellular Component: cell surface; external side of plasma membrane; extracellular region; extracellular space; integral to plasma membrane; lipid raft; phagocytic cup; plasma membrane; recycling endosome
Molecular Function: cytokine activity; identical protein binding; protease binding; protein binding; tumor necrosis factor receptor binding
Biological Process: activation of MAPK activity; activation of MAPKKK activity; activation of NF-kappaB transcription factor; caspase activation; chronic inflammatory response to antigenic stimulus; cortical actin cytoskeleton organization and biogenesis; embryonic gut development; I-kappaB kinase/NF-kappaB cascade; induction of apoptosis via death domain receptors; inflammatory response; leukocyte tethering or rolling; lipopolysaccharide-mediated signaling pathway; MAPKKK cascade; negative regulation of cytokine secretion during immune response; negative regulation of interleukin-6 production; negative regulation of lipid catabolic process; negative regulation of protein complex disassembly; negative regulation of transcription from RNA polymerase II promoter; negative regulation of transcription, DNA-dependent; negative regulation of viral genome replication; positive regulation of apoptosis; positive regulation of caspase activity; positive regulation of cell adhesion; positive regulation of chemokine biosynthetic process; positive regulation of chemokine production; positive regulation of cytokine production; positive regulation of cytokine secretion; positive regulation of fever; positive regulation of heterotypic cell-cell adhesion; positive regulation of I-kappaB kinase/NF-kappaB cascade; positive regulation of interleukin-8 biosynthetic process; positive regulation of interleukin-8 production; positive regulation of JNK activity; positive regulation of MAP kinase activity; positive regulation of membrane protein ectodomain proteolysis; positive regulation of NF-kappaB import into nucleus; positive regulation of NFAT protein import into nucleus; positive regulation of nitric oxide biosynthetic process; positive regulation of osteoclast differentiation; positive regulation of peptidyl-serine phosphorylation; positive regulation of phagocytosis; positive regulation of programmed cell death; positive regulation of protein amino acid phosphorylation; positive regulation of protein complex assembly; positive regulation of protein complex disassembly; positive regulation of protein kinase activity; positive regulation of protein transport; positive regulation of smooth muscle cell proliferation; positive regulation of transcription factor activity; positive regulation of transcription from RNA polymerase II promoter; positive regulation of transcription, DNA-dependent; protein import into nucleus, translocation; protein kinase B signaling cascade; receptor biosynthetic process; regulation of I-kappaB kinase/NF-kappaB cascade; regulation of insulin secretion; response to glucocorticoid stimulus; response to salt stress; response to virus; sequestering of triacylglycerol; tumor necrosis factor-mediated signaling pathway
Disease: Asthma, Susceptibility To; Malaria, Susceptibility To; Migraine With Or Without Aura, Susceptibility To, 1
Research Articles on TNF-alpha
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Product Notes
The TNF-alpha tnf (Catalog #AAA668892) is a Cell Line and is intended for research purposes only. The product is available for immediate purchase. AAA Biotech's TNF-alpha can be used in a range of immunoassay formats including, but not limited to, Functional Assay. Application: *Monitor the TNF-alpha induction activity. *Screen for activators or inhibitors of the TNF-alpha signaling pathway. Culture conditions: Cells should be grown at 37 degree C with 5% CO2 using DMEM medium (w/L-Glutamine, 4.5g/L Glucose and Sodium Pyruvate) supplemented with 10% heat-inactivated FBS and 1% Pen/Strep, plus 3 ?g/ml of Puromycin. It is recommended to quickly thaw the frozen cells upon receipt or from liquid nitrogen in a 37 degree C water-bath, transfer to a tube containing 10 ml of growth medium without Puromycin, spin down cells, resuspend cells in pre-warmed growth medium without Puromycin, transfer resuspended cells to T25 flask and culture in 37 degree C-CO2 incubator. Leave the T25 flask in the incubator for 2~4 days without disturbing or changing the medium until cells completely recover viability and become adherent. Once cells are over 90% adherent, remove growth medium and passage the cells through trypsinization and centrifugation. At first passage, switch to growth medium containing Puromycin. Cells should be split before they reach complete confluence. To passage the cells, detach cells from culture vessel with Trypsin/EDTA, add complete growth medium and transfer to a tube, spin down cells, resuspend cells and seed appropriate aliquots of cells suspension into new culture vessels. Subcultivation ration = 1:10 to 1:20 weekly. To achieve satisfactory results, cells should not be passaged over 16 times. Researchers should empirically determine the suitability of the TNF-alpha tnf for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "TNF-alpha, Cell Line" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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